Yak’s Biochemistry Contain the Factors that Make it Possible for Human Habitation on the Top of the World in Central Asia

Introduction

In English terminology, Yak is used for both sexes. In Himalayan terminology, Yak is used for the male and Dzomo or Zhom for the female (Figure 1).

Figure 1 : Wild Yak, Himalayas (Photographed by Lopa Basu).

Materials and Methods

Description

The yak have enlarged lungs (four times as large as cattle), and more numerous red blood cells (three times more than cattle) and smaller cells (1/2 the size of cattle). Yaks have a dense coat which may be red, brown or black. They also have long hair sometimes reaching the ground (kilts or skirt) and lower number of sweat glands for conserving heat. Strong solid horns are used for removing snow to obtain covered vegetation and also used to establish the pecking order. They have multiple stomachs ruminant for digesting cellulose and coarse grazing material. They are also divided into domesticated (smaller size) and wild (larger size) yak. When it gets too cold wild yaks migrate seasonally to the lower plains to eat grass, lichens’ (composite symbiotic organism of algae and/or cyanobacteria living among multiple fungi) and herbs.

When it gets too hot they retreat to the higher plateau. Unfortunately, the numbers of these animals are decreasing due to uncontrolled hunting, predators, diseases, decreasing grazing areas and quality, malnutrition combined with the unforgiving environment. Yak nicknames are “Grunting Ox” or Hairy Cattle. The domesticated animals are Bos Grunniens and the wild is Bos mutus and they are native to the Himalayan Mountains of China Tibet, Nepal, and the surrounding areas. They were domesticated over 3,000 years ago, and can cross with bison and cattle. The male hybrids are sterile but the females (Dzomo or Zhom) are fertile. Yaks have short legs (females 4.5 feet in height and males 5.5-6.5 feet) with a weight of 1,800- 2,200 pounds. If dairy cattle are used in the cross the milk production is increased but most owners prefer pure breeds. The yaks (both sexes) have large horns (30 inches), four compartment ruminant stomachs, and a shoulder hump. Yaks head droops before high massive shoulders. They are sure footedanimals for the rough terrain. Thick red, black or brown undercoats acts as insulation and keep them warm to -40 °F or C. Domesticated yaks are more variable in color, and white splotches are common.

Reproduction

Heifers will breed at 18 months of age and gestation period is 8.5 months and males are of breeding age when then they are three years old. Calves at birth weight about 30 pounds but grow quickly on mother’s milk but the average survival rate is only about 50% due to poor nutrition for both mother and calf, harsh environmental conditions and death due to predators and diseases. Wild yak reproduce every two years and domestic yak sometimes reproduce ever year and occasionally have twins if the mothers have adequate nutrition. Infant yak can live up to 20 years.

Social

Most wild yak cows live in large herds with their young in groups of up to 100 or more animals. In contrast, adult males spend most of the year alone or in small groups.

Uses Of Yak

Meat – from grass fed animals is usually very lean and the yak is no exception.

Hair – When yaks shed their red, blackish or brown undercoat in the spring, the hair can be collected, combed out, and processed and results in a fiber that is comparable to cashmere, angora or qiviut (inner wool of the muskox). The coarser brown or black outer hair or ‘guard hair’ is traditionally used to weave ropes, belts, and bags. Hides – Can be tanned and used for many leather products including tent material and leather clothing.

Horns and Bones – Are often carved into utensils or sold to tourists or shipped (often on this beast of burden) to more traveled areas. Bones also find domestic uses, are converted into jewelry or carved into art objectives. Horns are also carved into objects of art. Fuel- Dried dung of the yak is the only obtainable fuel in this area of the world and is sometime used as construction material.

Yak Meat

Yak meat is a staple and can be naturally frozen by the environment but drying is often used to increase shelf life. It is a sweet, juicy, ultra-lean dark red delicately flavored red meat that is not gamey. It is lighter tasting than beef and never greasy. Meat can be air-dried and will keep from one to two years. Smoked meat (bacon beef) requires one to two days in salt and then keeps for up to 2 years. Spiced jerky or curry jerky is also available. Vacuum packaging can reduce oxidation of yak meat during frozen storage.

Meat Composition

Only a few papers have addressed this topic but one of the more informant ones on yak meat is by XD Zi, GH Zhong [1-3]. The data in Figure 2 and Tables 1 & 2 is a summary from this source.

Other Uses

Beast of burden-Yak can transport salt, grain and etc. They are used for plowing and threshing grain (primarily barley). Some yaks can be saddled and ridden. Yak racing is also practiced along with yak skiin and yak polo. Milk – Yak cow milk (higher in butter fat than cows) makes excellent cheeses (chhurpi) and is often dried to increase shelf life. Butter which is consumed in large quantities is also used in lamps and made into butter sculptures used in religious festivities. Butter is a staple food with salt added. Sour buttermilk can also be added to milk to produce sour milk. Grain is sometimes added to milk which results in the main course meal. Milk-tea, a mixture of tea and milk is often served to guests and is yellow in color.

Milk and mushrooms are used to make a stew; salt is usually added but no additional sugar is needed since the milk is naturally sweet in flavor. Yogurt is produced which also contributes needed nutrition. Milk powder is just beginning to become popular. Renin is added to coagulate the milk to make cheese. Viscera (edible) includes heart, stomach plus ruminant, small and large intestine, liver, kidney. In the inedible category, lungs and pancreas are primarily used for dog feed even though they are considered edible in other areas of the world.

Climate

Yaks have been brought to warmer climates and placed in zoos and some can be found on ranches where their primary food is grass. Under these conditions their meat sells well since it is different and scarce but the animals seem to prefer a colder climate.

Result and Discussion

Shows the live weights of Yak at different ages. It is evident from this graph that both male and female yaks have similar birth weights but starting at age 2 and continuing through 8 years the males out gain the females and the final mature weight of the males is about twice as much as the females (Figure 2) and (Table 1). For dressing percentages, the bull and steer are not significantly different but the female has a significantly lower percentage. For back-fat thickness, comparing all the values the results indicate no significant difference due to sexes. Marbling scores indicate all sexes are significantly different with the steer having the highest amount followed by the bull and the female which had a significantly lowermarbling score. For color, the steer had a significantly higher color values followed by similar scores for both bull and female.

Figure 2 : Live weights of Yak at different ages.

Table 1 : Carcass characteristics of Yak.

For pH (1h), the steer and female had similar scores but the bull was significantly higher. For pH (24h), all three sexes were similar but they are higher than would be expected for cattle suggesting that this may have been genetics or the animals may have been stressed before they were harvested. Also in general grass fed animals normally have higher ultimate pH values than grain fed animals (Table 2).

Table 2 : Proximate Composition of 10th/12th rib-cut of Yak.

Proximate composition of 10th/12th rib-cut of Yak is shown in (Table 2). Present dry matter was significantly lower in bulls than in steers or females which were not significantly different. Percent protein was significantly lower in females than in bulls and steers which were not significantly different. Fat parentage was significantly lower in bulls than in steers or females which were not significantly different. Percent ash was not significantly different due to sex. Amino acids were also analyzed in the original research but values are not shown in this summarized version since most of the values were similar to the expected ranges. The only major difference was that methionine is dramatically lower (0.34 g/100 g protein).

Conclusion

Yaks, due to their genetic construction have the ability to prosper at high altitudes and withstand stand cold temperatures and sparse vegetation. Their existence makes human habitation possible under these same conditions.

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Bacteria A Benefit Cell

Mini Review

Before birth, microbiota does not exist in the human body, and microorganisms start to adapt in several locations in the body after birth. The microflora in the intestinal tract takes a long period to be formed. After they formed, human body get benefits from these microbiota and the important benefit the immune system supports, disposal of waste gastrointestinal, development metabolism and resistance the body against the pathogenic microorganisms [1-7].

Microbiota in the intestinal play a major benefit to the human health, the forming of these microbiota depended on the type of nutrients and lifestyle, and these reflected on the impact of formed microbiota on the body energy and metabolism [8-12]. Study the reflect of the food type on the intestinal microbiota were determined by several studies, López-Nicolâs et al. [13] studied the effect of fruit juices pine bark extract (PBE) on the intestinal microbiota and they found that the pure phenolic compounds such as gallic acid had a high antimicrobial effect on Staphylococcus aureus and Escherichia coli.

A study on the effects of lactose or inulin on constipated elderly patients of measured microbiota composition, the results show considerable interindividual variations that inulin increased bifidobacteria significantly and decreased enterococci in number and enterobacteria in frequency. Where the individuals consuming lactose, increase in fecal counts of enterococci and decrease in lactobacilli. Further, clostridia were detected. Total bacterial counts remained unchanged [14]. The dynamic mutual relations between the intestinal microflora and colon cancer risk can be modified by dietary components and eating behaviors. Composition of intestinal microbiota can be influenced by several dietary components.

Dietary modifiers the numbers and types of microbes and have been reduced colon cancer risk experimentally by generating bioactive compounds from food components. Further, gastrointestinal microbiota can impact both sides of the energy balance equation [15]. On the other hand, dieting affect the intestinal microbiota, several studies sought the relationship between the nutrient types and the composition of the intestinal microbiota, and the consequent of that on the human health and his activities. The determine of chronic soy consumption on modified gut microbiota activities, the results showed high concentrations of is flavones and their gut microbiota metabolites in the plasma, urine, and feces in the high-soy diet consumer compare to who consumed the low-soy diet [16].

In a study of dietary types to clarify some compounds etiologic role in colon cancer. The results showed that consuming a mixed Western diet was more able to hydrolyze glucuronide conjugates, and more microbially degraded bile acids compare with the vegetarians, seventh-day Adventists, Japanese and Chinese [17]. The probability of the effect of trans galacto oligo saccharides and a placebo on the composition and activity of the intestinal microbiota were determined in samples of males and females, and the concluded of this work confirmed that trans galacto oligo saccharides do not beneficially change the composition of the intestinal microflora, and After the ingestion of both placebo and trans galacto oligo saccharides number of bifidobacteria increased in intestinal microbiota [18].

Instead of the effect of the nutrient on the composition of microbiota, some factors had a side effect on the composition of macrobiotic. One of the most important factors is using the antibiotics; several studies investigate and determine the frequency and change of intestinal microbiota after a course of antibiotic treatment [19-21]. To study the effect of amoxicillin and vancomycin in the composition of microbiota randomized samples were measured to detect the changes in microbiota; the results showed that vancomycin reduced microbiota diversity and decrease gram-positive bacteria [22].

The antibiotics ciprofloxacin, clindamycin, vancomycin, amoxicillin, clarithromycin, and lansoprazole were studied as tream of long period antibiotic treatments. The conclusion of these studies improved the side effect of reduction and changes of the intestinal microbiota, facultative anaerobes and Enterobacteria increased after treatments with the antibiotics amoxicillin, clarithromycin, and lansoprazole, The influence of the intestinal microbiota differto the different antimicrobial agents in different ways, the effect in microbiota depends on several factors for example the agent spectrum, the dosage and duration of treatment [23- 25].

Moreover, treatment with chemotherapy has a side effect on the microbiota including changes in their compassion and their population, recently several studies investigated the effect of chemotherapy on the intestinal microbiota, 5-fluorouracil therapy infliance in methane produces by methanogenesis which reverse association with diarrhea and positive association with constipation, the intistenal microbitoa have been toxicity during the therapy [26].

Further, the impact of chemotherapy treatment on the composition of the microbiota extended to the composition of the human milk microbiota, a recent comparing study on the microbiota composition in cancer patient mothers and healthy mother’s milk showed that the genera Bifidobacterium, Eubacterium, Staphylococcus and Cloacibacterium were depleted, where the genera Acinetobacter, Xanthomonadaceae and Stenotrophomonas were decreased [27].

The gut microflora plays a major role to repair the intestinal homoeostasis and integrity. In a survey of intestinal microflora, bacteremia, mucositis, chemotherapy-induced diarrhea, chemotherapy-induced mucositis, radiotherapy-induced mucositis to evaluate the function of gut microflora in the pathogenesis of gastrointestinal mucositis, authors found that Patients who receiving cytotoxic and radiation therapy exhibit marked changes in intestinal microflora, with most frequently, decrease in Bifid bacterium, Clostridium cluster XIVa, Faecalibacterium prausnitzii, an increase in Enterobacteriaceae and Bacteroides, and they concluded that gut microflora can play a major role in the pathogenesis of mucositis by the changing of intestinal barrier function, innate immunity and intestinal repair mechanisms [28].

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Spontaneous Internal Drainage of Iatrogenic Oesophageal Perforation

Introduction

Oesophageal perforation remains a potentially fatal disease with mortality rates of 10% to 40% reported [1]. Iatrogenic injury is the most frequent cause of oesophageal perforation, with studies reporting incidence rates of 14% – 100% [2], followed by spontaneous rupture (Boerhaave syndrome) [3]. A variety of treatment options are available ranging from conservative medical therapy to radical oesophagectomy. Newer modalities with interventional endoscopy with or without placement of stents are more frequently being described [4].

Case Report

60-year old female with symptoms of dysphagia and reflux underwent diagnostic oesophago-gastroduodenoscopy (OGD). Immediately post-procedure, patient complained of chest pain and shortness of breath. Surgical emphysema was present in the neck and Computed Tomography (CT) confirmed oesophageal perforation in an upper oesophageal diverticulum (Figure 1). She remained well whilst managemened conservatively for 10 days with nasogastric tube, intravenous antibiotics and omeprazole. Subsequently the patient became septic, developing signs of mediastinitis and lung consolidation. She was transferred to Intensive care unit (ICU) where parenteral nutrition was commenced and antibiotics were changed appropriately. A repeat CT did not demonstrate any collections which required drainage, however there was a large iatrogenic oesophageal diverticulum extending from the retropharyngeal space down to the posterior mediastinum, just beyond the carina, compressing on the oesophagus. The patient also developed a neck swelling and air leak from a suspected perforated pharyngeal pouch for which ENT opinion was sought.

Figure 1:

Figure 2:

Repeat CT showed a small amount of contrast, identified within the gas containing mediastinal cavity which suggested a persistent small leak (Figure 2). The patient continued to be septic and did not improve as expected. Hence a repeat CT and contrast swallow was performed which demonstrated no leak in the cervical oesophagus, however there was a persistent defect. Contrast was noted to be leaking from a defect at the level of carina (Figure 3).

Figure 3:

A feeding jejunostomy was placed and a rigid OGD demonstrated a large defect in the thoracic oesophagus as well as perforated upper oesophageal diverticulum. ENT surgeons excised the diverticulum and drained the cavity. The distal perforation was left alone at this stage. This fortunately resolved spontaneously in 4 weeks without the need for further intervention. The upper oesophageal perforation/abscess cavity had internally drained itself back into the posterior thoracic oesophagus. Treating the proximal perforation led to spontaneous resolution of the distal perforation and abscess cavity.

Discussion

Flexible video endoscopy has almost totally replaced rigid oesophagoscopy. Despite the inherent safety of the procedure (0.03% perforation risk compared to 0.11% for rigid endoscopy), the dramatic increase in the number of examinations performed has led to an increase in the number of associated injuries [5]. Perforation related to diagnostic upper GI endoscopy occurs distally in 75 – 90%, mostly in relation to pathology. Most iatrogenic trauma is recognised immediately, or there is at least a high index of suspicion. Our patient presented soon after the procedure and appropriate management was commenced. However, this failed subsequently and she had to have multiple imaging only to diagnose a second oesophageal defect that was associated with an abscess cavity.

Iatrogenic cervical perforations are almost always contained and thus collections are usually drained percutaneously when necessary. Any resulting oesophagocutaneous fistulas heal rapidly in the absence of distal obstruction. Occasionally operative prevertebral lavage, primary closure and drainage is required, using a left lateral incision anterior to the sternocleidomastoid. This is well tolerated by even critically ill patients [5]. The optimal treatment of oesophageal perforation remains an issue of continuing debate. Surgical intervention has to be weighed against a conservative approach. Traditionally, the surgical approach was reserved to perforations diagnosed within a timeframe of 24 hr. If the diagnosis is not made early, surgery carries a high morbidity and mortality. Conservative therapy may be the better choice if the process is limited to the mediastinum, while surgery should be confined to those cases with simultaneous rupture or perforation of the pleura.

Our patient went onto develop a distal oesophageal defect thus internally draining the upper oesophageal perforation. Managing the upper oesophageal perforation lead to resolution and healing of the lower oesophageal defect. This is the first case to be reported in literature to clearly demonstrate that internal drainage and dealing with the proximal perforation could be attempted before contemplating thoracotomy if such a case is encountered in future.

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Syringomyelia in the Thoracolumbar Spinal Cord of an African Wild Dog (Lycaon Pictus)

Brief Communication

A 31-day-old, captive born, male, African wild dog (Lycaon pictus, AWD) displayed hind limb ataxia and weakness since the onset of ambulation. Separation of the animal carried a high risk of aggression to the litter from the pack, leading to a conservative medical approach throughout the case. At 36 days of age, the clinical signs became more notable and the animal was visually smaller and thinner than its littermates. Although able to stand, it displayed an unsteady gait, taking a few steps at a time before the hind limbs collapsed. On physical examination, the animal was bright, alert and responsive, had a body condition score of 2/5, and displayed postural reaction (PR) deficits in both hind legs, more pronounced on the left. The remainder of the neurological examination, including the thoracic limb reflexes and cranial nerve evaluation, was normal. Ulcerated skin calluses of 1 cm diameter were present bilaterally on the elbows as a result of the hind limb weakness and increased time spent in recumbency. Mild laxity in the coxofemoral joints was noted on palpation. Radiographs showed left coxofemoral joint subluxation and mild muscle atrophy of the left hind limb. There was also asymmetry of the left capital femoral physis. A computed tomography (CT) scan was performed (HiSpeed CT/I, GE Medical Systems, Waukesha, Wisconsin 53188, USA; 3mm slice thickness, 1.25 mm reconstruction interval, soft tissue reconstruction algorithm) that supported the findings of the left hind limb muscle atrophy, but could not confirm the coxofemoral joint changes seen on radiographs. No abnormalities of the vertebral column or spinal cord were detected. The gait abnormalities were thought to be due to laxity in the tendons and ligaments of the coxofemoral joints.

As the animal grew, its ability to ambulate improved, but the gait remained abnormal with the animal intermittently falling to the side with both hind limbs. At 45 days of age it was able to keep up with the pack and by 52 days the elbow lesions were resolving. At 66 days of age, the animal was reevaluated. Body condition score had declined to 1.5/5 and new superficial abrasions were present on the dorsal lateral aspect of both hind feet, due to its abnormal ambulation. Significant muscle atrophy and marked ataxia in the hind end were noted. The animal was able to rise to a standing position and ambulate, but would drag its hind legs more often than previously observed, especially at a run. PR was now completely absent in both pelvic limbs. No pain was elicited on spinal or neck palpation, and deep pain was present bilaterally in the hind feet. Mentation was still normal, thoracic reflexes were normal, and anal tone was present. The animal was suspected to have a spinal lesion caudal to third thoracic vertebrae (T3), given a guarded prognosis, and returned to the litter with close monitoring of quality of life.

The gait abnormalities progressively worsened as the littermates were maturing and integrating into the pack. On day 87 the animal sustained a bite wound to the cranium, prompting intervention for treatment and further assessment. Cranial nerve examination and thoracic limb reflexes were normal, and mentation remained appropriate, but the animal was now paraparetic. Multiple superficial abrasions were noted on the elbows, hocks, and feet. Cerebrospinal fluid (CSF) was collected from the L5-6 intervertebral space. Culture revealed no growth of aerobic or anaerobic organisms. Fluid analysis showed a mild mononuclear pleocytosis (total nucleated cell count (TNCC) = 5 cells/uL, 58% large mildly vacuolated mononuclear cells, 42% small lymphocytes), a mild increase in protein (61.7 mg/dL), and peripheral blood contamination (586 red blood cells (RBC)/uL). Normal canine reference intervals for this laboratory include: TNCC <5 cells/uL, protein concentration <45 mg/dL for lumbar samples, and >500 RBCs/uL was considered peripheral blood contamination. Findings were considered non-specific and typical of inflammation, infection, or trauma to the central nervous system.

Figure 1 : Sagittal T2-weighted magnetic resonance image showing hyperintensity within the spinal cord representing syrinx formation at the level of T13-L1 (arrow).

A magnetic resonance imaging (MRI) scan (Philips Gyroscan NT, Imaging Center for Animals, Buffalo Grove, IL 60089, USA) showed a non-contrast enhancing, T2-weighted hyperintensity within the central and dorsal aspect of the right spinal cord extending from the level of the caudal aspect of T13 caudally to the level of L1-L2 (Figure 1). There was also mild bulging of the L7-S1 intervertebral disc without significant compression. The lesion visualized via MRI and the progressive worsening of clinical signs support the diagnosis of syringomyelia of the thoracolumbar spinal cord. Surgical intervention was not pursued due to the extensive postoperative care and aggressive social behavior of AWD. Mounting concern for the pack injuring or killing this debilitated member prompted euthanasia.

Figure 2 : (A) Histological section, spinal cord, T13. The image is not the most affected portion of the spinal cord, but demonstrates the syrinx clearly. The right dorsal and lateral funiculi contain a well-demarcated cavitation, or syrinx, delineated by mildly compressed neuropil (arrow). The syrinx does not communicate with the central canal. The central canal is mildly ectatic. “R” indicates right side. Hematoxylin and Eosin. (B) Gross spinal cord, T13-L1. Multiple cross sections of T13 to L1 spinal cord have a variable sized cavitation within the gray and white matter. This cavitation is 1.5 cm long and up to 3mm diameter. Fixed sections (10% buffered formalin).

On necropsy, the animal had adequate subcutaneous and visceral adipose tissue stores. Epaxial, gluteal and hind limb skeletal muscles were markedly atrophied. Bilaterally, skin overlying the olecranon processes of the elbows was ulcerated and overlain by a serocellular crust, interpreted as pressure sores. A 1.5 cm long, up to 3 mm diameter round to irregular cavitation, or syrinx, in the T13-L1 spinal cord was identified that markedly compressed and distorted the adjacent neuropil. On histopathology, the T13-L1 spinal cord had a well-demarcated cavitation, or syrinx, involving the dorsal, lateral and ventral funiculi white matter as well as adjacent grey matter (Figure 2). The syrinx was circumferentially demarcated by mildly compressed neuropil. Adjacent axon sheaths were dilated and empty, or contained swollen spheroids, which indicate axonal loss and degeneration, respectively. A few empty axon sheaths contained a single gitter cell. The bulging disc noted via MRI was not appreciated grossly. The necropsy findings confirmed the diagnosis of syringomyelia.

Syringomyelia is a general term that characterizes a spinal cord cavitation containing fluid [1]. Syringomyelia is a secondary finding due to a primary cause. A cavitation in the spinal cord, or a syrinx, has been associated with primary disorders including spinal cord tumors, myelitis, meningitis, congenital vascular malformations, chronic arachnoiditis, and spinal cord trauma [1]. The cause of syringomyelia is unknown, but may result from venous obstruction, mechanical disruption, or shearing of spinal cord tissue planes.

Clinical signs associated with syringomyelia depend on lesion location within the spinal cord [2]. In this case, the hind limb paresis, weakness, and lack of normal postural reactions of the pelvic limbs, yet normal forelimb reflexes were consistent with deficits in the sensory and/or motor components of the central nervous system, and localized the lesion to the T3-L3 spinal cord segment [3]. While the diagnosis of the large syringomyelia at the T13-L1 spinal cord was deemed the cause of the gait abnormalities, the size of the syrinx determined by MRI does not always correlate with severity of clinical signs [4]. Cavitation lesions may be progressive and expand along the gray matter of the dorsal horns, which are planes of structural weakness, resulting in subsequent necrosis and edema of the spinal cord parenchyma around the cavitation and progression of clinical signs [2]. In this case, clinical signs progressed as hind limb PR was delayed at one month of age and absent by three months of age.

Syringomyelia has been documented in humans [4], domestic dogs [2,5,6], domestic cats [7], cattle [8], horses [9], an African lion [10], and a dromedary camel [11]. To the authors’ knowledge, this is the first report of syringomyelia in any non-domestic canid. Previously thought to be a rare condition, diagnosis of syringomyelia in domestic companion animals is now more common, especially with the increased availability of MRI. This disorder is well described in certain dog breeds, especially the Cavalier King Charles spaniel [1]. In dogs, Chiari-like malformations are the most commonly described cause of syringomyelia, which typically occurs at the craniocervical junction. Syringomyelia caudal to the cervical spinal cord has been associated with spinal dysraphism [5], vertebral malformations, intervertebral disc disease and arachnoid cysts [6].

Diagnosis of syringomyelia is commonly made via breed disposition, clinical signs, MRI findings, and CSF analysis. When syringomyelia is detected via MRI, the primary cause should be sought out both cranial and caudal to the lesion [12]. MRI in this case showed a mild bulging of the L7-S1 intervertebral disc, caudal to the spinal lesion. The intramedulary pulse pressure theory suggests syringomyelia can develop a distance from the obstruction of CSF flow in any part of the spinal cord [1], and a bulging disc could have narrowed the diameter of the vertebral canal enough to alter the CSF fluid dynamics. The MRI did not show significant compression of the spinal cord from the bulging L7-S1 intervertebral disc, making it a likely incidental finding and not the cause of the syrinx formation.

While MRI is the best diagnostic tool, collection of CSF can indicate the degree of the spinal cord involvement. In a study of dogs diagnosed with Chiari-like malformations, CSF of dogs with syringomyelia had higher total nucleated cell count, higher protein concentration, and an increased neutrophil percentage. There was also a strong positive correlation between total nucleated cell count and the size of the syrinx [13]. Disruption of the blood-spinal cord barrier resulting in increased permeability has been suggested as the most likely mechanism of increased CSF protein concentration with syringomyelia. In this case, there was a mild increase in total nucleated cells and protein concentration.

Medical management of syringomyelia is directed towards analgesia, corticosteroids, and drugs that decrease CSF production. Surgical intervention is indicated when neurologic deficits are present, with the intent of restoring CSF dynamics by addressing the primary cause [6]. Direct draining of the syrinx with stents or shunts does not carry a good long term outcome in people due to obstructions [1]. Surgery was indicated for this AWD based on the severity of the clinical signs, but the extensive post-operative care would have prevented re-introduction to the pack and was not deemed in the animal’s best welfare.

Syringomyelia associated with Chiari-like malformations in Cavalier King Charles spaniels is a complex trait that has a moderately high heritability [14]. Although the syrinx that occurred in this AWD was not due to a Chiari-like malformation, it is possible that the primary malformation is heritable. Decreasing genetic diversity is expected to increase the frequency of congenital and heritable diseases. The AWD population is endangered and has experienced a significant loss of genetic diversity in the managed population, which is contributing to development of a genetic bottleneck for this species [15]. At present, there is insufficient evidence from this single case to suggest syringomyelia is a heritable condition in this species, but this should be closely monitored in populations under professional care. This report highlights the importance of species specific breeding management, and the need for careful planning to maximize the genetic diversity and minimize potential of genetically heritable diseases.

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New Insight in Human Lactonase PON2

Mini Review

It is well established that oxidative stress from mitochondria plays an important role in apoptosis and also leads to premature aging and cancer [1]. There is growing scientific consensus that proteins with antioxidative functions, such as paraoxonases, can lower the incidence of these diseases. The paraoxonase (PON) gene family consists of three members: PON1, PON2, and PON3. PON genes are located in a gene cluster on chromosome 7q21.3-22.1 in human [2] and show about 70% sequence identity among them [3]. PON2 is a calcium-dependent glycoprotein of about 44 kDa, expressed ubiquitously, and associated with plasma membrane fractions. Recently, it has been demonstrated that PON2 is a type II Tran membrane protein, with its N-terminal region identified as a single Transmembrane domain, whereas the catalytic domain, corresponds to the C-terminus, located extracellularly to counteract lipid per oxidation [4].

PON2 has two main activities: a calcium-dependent hydrolytic activity, involved mainly in the hydrolysis of lactones, esters and aryl esters [2] and a redox function, which reduces the levels of ROS (reactive oxygen species) thus curbing cell oxidative stress and therefore displaying an anti-apoptotic effect. No clues are available about the molecular basis of this anti-ROS activity except the demonstration that PON2 can bind ubiquinone (coenzyme Q10) with high affinity only in the presence of Ca2+ [5]. In doing so coenzyme Q10 is substracted from interaction with O2 and production of ROS.

Unlike PON1 and PON3, PON2 is an intracellular protein, expressed in a wide range of cell types, including pancreatic beta cells. Importantly, polymorphisms in the PON2 protein (S311C and A148G) have been associated by several studies with diabetes and its complications. Although a genetic association between PON2 and these conditions has been shown, the exact function of PON2 in humans is not known. It has been shown that all of the PONs can inactivate the lactone 3-Oxo-C12-HSL [2,6-8]. Among PONs, PON2 has the greatest lactonase activity against the bacterial quorumsensing (QS) molecules [2,9]. QS Is an interbacterial mode of communication accomplished through the coordinated production, secretion, and detection of chemical signals (QS signals) that trigger the expression of specific bacterial genes. The QS signals self-produced by P. aeruginosa are in the form of small molecules, termed acyl- homoserine lactones (acyl-HSLs) [10] among which 3-Oxo-C12-HSL is the master regulator [11]. It has been demonstrated that PON2 responds to the bacterial quorormone 3-Oxo-C12-acyl homoserine lactone by a rapid decrease of the lactonase activity via a putative post-translational modification (PTM) [12]. Therefore, Pseudomonas produced 3-Oxo-C12-HSL, organizes from one side the coordinated attack of bacterial cells against human tissues and on the other curbs the human defences by blocking PON2 activity, which is thought to represent the first line of defense against infections [9].

PON2 being located on cell surface [4] can rapidly clear out the 3-Oxo-C12-HSL accumulating outside the cell and passing through the membrane. The increase in the concentration of the quorormone activates, at a certain threshold, the production of virulence factors and the formation of a biofilm [11]. It also diffuses inside the cells or binds specific receptors thereby activating processes that are different in different cell types [13]. Relevant is the effect on NFkB signaling through which 3-Oxo-C12-HSL attenuates the innate immune system to establish and maintain local persistent infection in humans, for example, in cystic fibrosis patients [14].

Recently we discovered a new PTM of PON2 induced by 3-Oxo-C12-HSL [15] namely the ubiquitination of Lys 144 (168 in the paper) that may be of physiological importance because it decreases the PON2 activity. Moreover we have shown that PON2 is able to interfere efficiently with the formation of the biofilm in vitro [15]. These experiments were conducted by expressing an engineered version of PON2 in E. coli and renaturing it from inclusion bodies. We also suggested in this way that glycosylation of PON2 is not important for activity but likely for stability. Teiber [16] reported that phosphorylation of Ser36 is necessary for activity. This is in contrast with our data: our rPON2 made in E. coli is active and devoid of phosphorylation also because the deletion at the N-terminus includes Ser36. In any case 3-Oxo-C12-HSL does not seem to act via this phosphorylation site [16].

In previously published large scale mass experiments Tyr346 has been found phosphorylated [17]. Mutants at that position maintain activity and are still inactivated by 3-Oxo-C12-HSL [16]. In our effort to clarify the role of PON2 we are focusing also on expression regulation. In a follows up of our previous work [18] we identified in the 3’UTRs of some MHCII genes a quite well conserved sequence, already suggested in a recent paper as a putative regulative signal [19]. The sequence was shared by a cluster of a limited group of genes. In the same database we looked for the presence of signals downstream of the PON2 gene. There was just one occurrence for PON2. The identified sequence was a 12mer: TTTCCTTAAAAT identifying a cluster of 21 genes. This sequence does not overlap with any MREs. The interesting aspect of this finding is that most of these genes are functionally related (based on information at http://www.genecards.org) and involved in apoptosis, proteosomal degradation (ubiquitinylation) and DNA repair (Table 1). We are currently analysing the involvement of these genes in regulation and physiology of PON2.

Table 1:

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Mineral Contents of Selected Medicinal and Stimulating Plants in Ethiopia

Medicinal Plants

Thyme

Thyme is cultivated in almost every country, as an aromatic for culinary uses. The two species, Thymus schimperi Ronniger and Thymus serrulatus Hochst. ex Benth are endemic to the Ethiopian highlands growing on edges of roads, in open grassland, on bare rocks and on slopes, between 2,200-4,000 m altitudes. Both species are perennial herbs, woody at the base and 5-40 cm high. The volatile oil from thyme was found to contain p-cymene, γ-terpine, carvacrol, rosmarinic acid, eugenol, and thymol [1]. The volatile oil not only has carminative action, but also antiseptic, antimicrobial and antifungal activities [2]. In the Ethiopian traditional medicine the plant has many medicinal applications. Some of the reported applications are for the treatment of gonorrhea, cough, inflammation, spasm, thrombosis, urinary retention, mental illness, eye disease, toothache, stomach problems, leprosy, lung TB, acne, and ascaris [3].

The fresh or dried leaves of both species are used locally as condiments in the preparation of chili powder, stew, bread and tea. The contents of some selected metals Ca, Mg, Fe, Mn, Co, Cu, Zn, Ni, and Cd in different thyme leaf samples widely consumed in Ethiopia were determined by flame atomic absorption spectrometry. The level of the minerals in the four samples ranged from 1,239-2,517, Ca; 1,524-1,786, Mg; 728-2,517, Fe; 37.7-114, Mn; 2.59-4.3, Co; 7.69-9.3, Cu; 8.7-52, Zn; and 9.83-14.2 μg/g, Ni; respectively. While the level of toxic metal Cd in the four samples ranged from 0.87- 1.3 μg/g. The concentration of Ca was higher than the other metals in the three samples and Cd was the least of all the metals in the analyzed samples [4].

Gesho

(Rhamnus prinoides) (Amharic, Gesho) is a wide spread plant species in Ethiopia and other east and south African countries. R. prinoides has been used for bitterness in the traditional brewing process in East Africa. R. prinoides has potential use as a commercial hopping agent in the beer industry. An extract of leaves and stems of this plant (a naphthalene glycoside, Geshoidin) is responsible for bitterness in local alcoholic beverages [5]. In Ethiopia, the leaves and stems of this plant are used to impart the characteristic bitter flavor to domestically brewed beverages such as Tella, Tej [6]. R. prinoides has also several traditional medicinal uses in Africa. A decoction of the root is taken as a blood purifier, to treat pneumonia, gonorrhea, rheumatism, stomach-ache, flu/ cold, back pain, brucellosis, strength/nutrient supplement, and enhancing digestion [7]. The boiled soup of the root is used for the treatment of common cold, leaves are used for chest pain and leaves/stems are used for the treatment of tonsil in central Kenya [7].

Extract of R. prinoides is used for the inhibition of Alzheimer’s disease. Extract of the leaf and root bark of R. prinoides has been also used for malaria treatment either alone or in combination with chloroquine in Kenya. The levels of essential metals (Ca, Mg, Cr, Mn, Fe, Co, Ni, Cu and Zn) and toxic metals (Cd and Pb) in the leaves and stems of Rhamnus prinoides (Gesho) cultivated in Ethiopia were determined by flame atomic absorption spectrometry. The levels (mg/kg) of the metals were found to be: Ca (6304-22236), Mg (3202-5706), Cr (5.08-20.6), Mn (8.12-17.9), Fe (47.9-187), Co (22.2-42.1), Ni (12.8-27.3), Cu (6.5-73.0), Zn (12.2-43), Cd (0.81- 3.10), and Pb (17.7-25.0) in the leaf samples and Ca (3601-5675), Mg (2635-5528), Cr (ND-16.3), Mn (2.16-3.98), Fe (22.0-124), Co (18.7-91.7), Ni (9.68-19.2), Cu (16.8-233), Zn (17.4-28.2), and Cd (ND-1.56) in the stem samples. The study showed that the leaf and stem of (R. prinoides) (Gesho) are good sources of essential minerals and virtually free from toxic metal Cd but not from Pb in the leaf [8].

Linseed

Linseed (Linum usitatissimum) stands fourth after mustard, sesame and groundnut in edible oil production of the world. Linseed has been a traditional crop in Ethiopia. Ethiopia is the 5th major producer of linseed in the world after Canada, China, United States and India [9]. Linseed is a multi-purpose crop. Its’ seeds containing about 36-40% of oil, have long been used in human and animal diets and in industry as a source of oil and as a basic component or additive of various paints or polymers. Recently, there has been a growing interest in the probiotic properties of flax and in its beneficial effects on coronary heart disease, some kinds of cancer and neurological and hormonal disorders [10]. The beneficial effects are mostly due to flax lipids. Flax oil is the richest plant source of linoleic (omega-6) and linolenic (omega-3) polyunsaturated fatty acids, which are essential for humans since they cannot be synthesized in the organism and must be ingested in food [11].

Scientific research over the past decade all over the world is indicating health benefits of omega-3 type oils, lignin and other soluble fiber present in the flax seed/linseed. Some of the medicinal uses of linseed oil are: flax seed oil mixed with an equal quantity of limewater, is an excellent applicant for burns and scolds, rectal injection has been recommended for piles, freshly extracted oil is used as a laxative, linseed oil is a vehicle for irritant drugs, linseed tea is used as a demulcent in cough especially those forms due to irritation of pharynx and upper part of respiratory passage. It is also used as a demulcent drink in intestinal or urinary catarrhs), for the preparation of cough syrup mucilage of linseed is used, crushed linseed is used in the form of poultice to apply warmth and moisture locally for the relief of superficial or deep rooted inflammation. The poultice may be sprinkled with boric acid previous to application.

The poultice mass is enclosed in muslin, the surface of poultice may be smeared with oil to keep it adhering to skin), linseed/flax seed and its oil have anti-inflammatory action the treatment of arthritis, feeding linseed and its oil may be indicated in hyperlipidemia to decrease platelet aggregation and also to reduce and control atherosclerosis, thrombosis and myocardial infarction by reducing cholesterol and low density lipids. The levels of essential and non-essential metals in linseed samples collected from five different sites (Bale, East Gojam, Shoa, South Wello and Tigray) in Ethiopia were determined by flame atomic absorption spectrometry. The levels (mg kg^-1) of metals determined were in the ranges Na (242-614), K (6,494-6,755), Mg (2,679-3,118), Ca (540-744), Cr (13-30), Mn (17-28), Fe (198-242), Co (23-42), Ni (12-16), Cu (25-45), Zn (29-40), and Pb (12-32). Cd was not detected. K and Fe were with the highest concentration from major and trace metals, respectively. The study indicated that Ethiopian linseed is a good source of essential metals and free from the toxic metal Cd but not from Pb [12].

Croton

Ethiopia has a long history of traditional medicine and has developed ways to combat disease through it. In Ethiopia up to 80% of the population uses traditional medicine due to the cultural acceptability of healers and local pharmacopeias, the relatively low cost of traditional medicine and difficult access to modern health facilities [13]. Of the many medicinal plants in Ethiopia, Croton macrostachyus is the most common. Croton macrostachyus Hochst. ex Del. is commonly known as rush foil or broad-leaved Croton. It is native to Eritrea, Ethiopia, Kenya, Tanzania, Uganda and Nigeria. As C. macrostachyus is available almost in all parts of Ethiopia, people are using it as a medicine for treatment of snakebite, malaria, headache, internal worms, rabies, gonorrhea, ascarasis, sexually transmitted diseases and tinea versicolor for human being in which the traditional medician ordered the patients to take unlimited dosage of the plants part like to take two, three or even more glasses of medicinal plant extract orally.

The plant is also used for fever and wounds of domestic animals [14]. The leaves of Croton macrostachyus have medicinal value including cold leaf decoction is drunk or ashes taken orally as treatment for cough; juice from fresh leaves is applied on wounds to hasten clotting. The levels of essential metals (Ca, Mg, Fe, Mn, Zn, Cu, Co, Cr, Ni) and toxic metals (Cd, Pb) were determined in the leaves of Croton macrostachyus (traditional medicinal plant) collected from four different regions of Ethiopia (Akaki, Abomsa, Bonga and Dilla) and also in the infusions of leaves collected from Akaki using flame atomic absorption spectrometry. The mean concentration ranges (μg/g) were Ca (5,823-12,040), Mg (1,971-4,961), Fe (192-581), Mn (157-1,770), Zn (19.5-60.5), Cu (6.31-18.6), Co (1.97-3.45), Cr (2.13-8.75), Ni (2.15-3.80), Cd (0.75-1.08) and Pb (1.05-2.19) in the leave powders and Ca (716-1,776), Mg (16.7-80.9), Fe (1.39-3.34), Mn (2.17-3.40), Zn (0.674-7.88), Cu (1.94-2.31), Co (0.157-0.224), Cr (0.144-1.23), Ni (0.203-0.267), Cd (0.05-0.098) and Pb (0.0148- 0.185) in the infusion samples. Among the quantified metals in the infusion samples, Ca showed the highest leaching rate (61.5%) in the 24 h infusion while Fe exhibit the lowest (1%) leaching rate in the 3 h infusion [15].

Fenugreek

Trigonella foenum-graecum (fenugreek) is native to Ethiopia and to the area from the Eastern Mediterranean to Central Asia, and is widely cultivated in Pakistan, India and China [16]. Fenugreek seeds contain about 50% fiber and also contain protein, saponins and the hypoglycemic phytochemicals coumarin, fenugreekine, nicotinic acid, phytic acid, scopoletin and trigonelline and 4-hydroxyisoleucine. Fenugreek seeds have high contents of iron, calcium, zinc, lysins and β-carotene and are a rich source of flavonoid compounds such as quercetin, luteolin, kaempferol, tricin and gallic acid [17]. Fenugreek leaves and seeds are consumed in different countries around the world for different purposes, such as medicinal uses (anti-diabetic, lowering the blood sugar and cholesterol levels, anti-cancer, anti-microbial, against breast cancer, avoiding blood poisoning from wounds, making food (stew with rice in Iran, flavoring cheese in Switzerland, syrup and bitter rum in Germany, mixed seed powder with flour for making flat bread in Egypt, curries, dyes, young seedlings for eating as a vegetable), roasted grain as a coffee-substitute (in Africa), controlling insects in grain storages and perfume industries.

The seeds of this ancient herb have been used as both a spice and a herbal remedy in the Middle East, India, and Egypt and later on in Europe, China and other parts of the world [18]. Fenugreek seeds have been reported for their pharmaceutical properties in treating such human diseases as diabetes and hypercholesterolemia [18]. In Ethiopia fenugreek is cultivated between altitudes of 1600 and 2300 m above sea level all over the country and is used in preparing “Hilbet”, a delicious, traditional, soft white food in the Tigray regional state. It is also used to make tea, “Enjera”, flat bread mostly made of “tef” flour, as a spice in milk and in traditional medicine. The levels of the major (Ca, K, Na, Mg), trace (Fe, Cr, Ni, Zn, Mn, Cu, Co), and toxic (Pb, Cd) metals in the seeds of fenugreek cultivated in different regions of Ethiopia were determined by flame atomic absorption spectrophotometry. Thirteen elements were determined, obtaining concentrations (mg kg^-1) in the following ranges: Ca (15353-36771) > Fe (6041-18584) ≈ K (6789-11517) > Pb (615-2624) > Na (201-1559) > Cd (285-464) > Cr (3-552) > Ni (31-108) > Mg (31-102) > Zn (15-33) > Mn (16-28) > Cu (ND- 35) > Co (4-15). The study showed that fenugreek seeds were a good source of essential metals. However, they also contained large amounts of the toxic metals Cd and Pb and therefore should not be consumed daily [19].

Stimulating Plants

Khat

Khat (Catha edulis Forsk) is an evergreen shrub or tree found growing wild or cultivated in the east of a region extending from Southern Africa to the Arabian Peninsula more specifically in Yemen, Ethiopia, Kenya, Madagascar, Somalia, Tanzania, and others as well. The most favored part of the plant is its leaves, particularly the young shoots near the top of the plant. However, leaves and stems at the middle and lower sections are also used. Khat is chewed for its stimulating property. This is due to the presence of the phenylalkylamines in the plant [20]. There is an ever-growing demand of khat in Ethiopia both for domestic consumption and for the export market. Most of the exported khat is grown in the eastern part of the country and mainly exported to the neighboring and the Middle East countries, and in recent years, the market for khat has grown to Europe and America [21].

Report indicates that over 20 million people in the Arabian Peninsula and East Africa chewed this plant daily [20]. Khat use is widespread and cultivated in most parts of Ethiopia, where its use is socially sanctioned and even prestigious [22]. In Ethiopia, the plant is marked under different names: Awadai, Kuto, Gelemso, Gurage, Wendo, Sebeta, Bahir Dar, Liyu, Chengie, Berdaye, Anferara, Colombia, etc. Of these, only some of them are commonly available in the capital city, Addis Ababa, and exported to the neighboring countries while the remaining is chewed by the local people around. The levels of essential (Ca, Mg, Mn, Fe, Zn, Cr, Cu, and Co) and nonessential (Cd and Pb) metal in six different varieties of Ethiopian khat chewing leaves were determined by flame atomic absorption spectrometry. The concentrations ranges in fresh-weight basis were recorded in decreasing order: Ca (1,038-2,173 μg/g) > Mg (478.2-812.3 μg/g) > Fe (53.95-82.83 μg/g) > Zn (5.18-9.40 μg/g) > Mn (6.98-8.66 μg/g) > Cu (1.85-5.53 μg/g) > Cr (0.66-3.47 μg/g) > Co (0.41-0.80 μg/g). A wide variation in the mineral contents of khat from different region of Ethiopia was noticed. The toxic metals (Pb and Cd) were not detected in all the samples analyzed [23].

Cannabis

Cannabis (Cannabis sativa L.) is an annual herbaceous plant. It is a dioecious plant. In many countries, cannabis is cultivated as a narcotic substance or a source of narcotic substances like hashish and hashish oil. Now a day’s cannabis is cultivated on the large areas with the mild and tropical climate for the cannabis oil and fiber [24]. The highest levels of cannabis production in the world take place in the African continent. The crude drug can be obtained from leaves, flowers, seeds and stem of cannabis. It can be smoked in cigarettes or pipes and can be snuffed or added to food [25]. Cannabis products are the most widely trafficked drugs worldwide. Practically all countries in the world are affected by cannabis trafficking [24].

A small amount of cannabis is produced in rural areas of Ethiopia, of which a small portion is exported, primarily to the neighboring countries; the majority is consumed at home, but absolute quantities in both cases are moderate. The chemical composition of cannabis varies with the type, age and part (flower, root, leaf, fiber, etc.) of cannabis plant as well as with the type of preparation. While Δ9-tetrahydrocannabinol is responsible for psychoactive properties of cannabis some of the other components modulate its activity. Cannabis sativa L. is known for millennia for its therapeutic properties and as a recreational psychoactive drug [26]. The levels of selected metals in leaves of Cannabis sativa L. cultivated in four different regions of Ethiopia were determined by flame atomic absorption spectrometry. The levels of metals determined (μg/g dry weight) were in the ranges Ca (657-1,511), Zn (321-380), Ni (124-172), Cu (122-176), Cd (3-10), Pb (8-10), and Cr (4-8). Zn was with the highest concentration among trace metals [27]. The results indicated that the content of Pb and Cd exceeded the permissible amount for medicinal plants which form the raw materials for the finished products set by World Health Organization (WHO) [27].

Tobacco

Tobacco (Nicotiana tabacum L.) is a commercial plant. Tobacco is one of the basic agricultural products, in Ethiopia, with social and economic importance. Native tobacco (‘gaya’) has been used for snuffing, chewing and for pipe smoking in many places. Three main types of commercial tobacco are produced in Ethiopia: Virginia,Oriental and Burley. Virginia accounts for a little more than 74% of the total production, followed by Oriental, 22%, and Burley, 4% [28]. United Nations Office on Drugs and Crime report [29] reveals that in Ethiopia peoples commonly use khat, tobacco, and alcohol, which have a share of 48.2, 29.9 and 18.9%, respectively, of all type of drugs. This clearly shows that tobacco made significant contribution as a drug in Ethiopia. Tobacco leaves are used for cigarette production and chewing. It naturally accumulates and concentrates relatively high levels of heavy metals and particular cadmium in leaves [30].

Cadmium is a non-essential to both plant and human. It is highly toxic and gets accumulated by tobacco plants. Most heavy metals cause a significantly serious damage on human health [31]. One of the main sources of toxic metals in our environment is tobacco smoke. Cigarette smoking is a major source of intake of these toxic elements not only to the smoker but also, through passive smoking, to nonsmokers. The distribution and accumulation of metals in tobacco leaves are the reflection of the mineral composition of the soil and environment in which the tobacco plant grows. Therefore, the actual metals content of tobacco vary considerably according geographic origin, the use of fertilizers with different chemical compositions and other characterizing features such as water for irrigation [32].

The levels of heavy metals (Cd, Cr, Cu, Ni, Pb and Zn) were determined in the raw and processed Ethiopian Virginia tobacco leaves collected from two different regions (Billate and Shewa Robit) of Ethiopia and three processed tobacco samples collected from National Tobacco Enterprise, Addis Ababa, Ethiopia by flame atomic absorption spectrometry. The mean metals concentrations (in μg/g dry weight) in the raw Virginia tobacco leaves from Billate and Shewa Robit, respectively, were: Cu (4.38, 7.30), Zn (53.7, 33.2), Cd (1.20, 1.30), Cr (ND, 1.45), Ni (ND, 1.90). The mean metals concentrations (in μg/g dry weight) in the processed tobacco from Billate and Shewa Robit, respectively, were: Cu (9.80, 12.8), Ni (2.35, 2.20) Cd (1.45, 1.90), Cr (1.65, 1.75), Zn (101, 83.8). The mean metals concentrations (in μg/g dry weight) in the processed tobacco Nyala (Ethiopian cigarette leaves) were: Cu (8.95), Cd (1.55), Cr (1.62), Ni (4.70), Zn (79.3). The study showed that the metal contents of tobacco leaves varied with the geographical origin in which the tobacco plant grows. The metal contents of processed tobacco were higher than the corresponding raw leaves. Pb was not detected in both the raw and processed Ethiopian tobacco leaves [33].

Conclusion

The level of metals in Ethiopian thyme is a comparable result with other medicinal plants. The concentration of Fe in thyme is higher than the values reported by different authors in other medicinal plants. The concentration of potentially toxic metal (Cd) in thyme ranges from 0.87-1.3 μg/g. This concentration is above the WHO acceptable level (maximum 0.3 mg/kg). The leaf and stem of (R. prinoides) (Gesho), which are used in the preparation of traditional alcoholic beverages in Ethiopia, are good sources of essential minerals and virtually free from toxic metals. The levels of metals in R. prinoides are in good agreement with the most of reported values for all the studied metals except Mn and Fe which are lower in the leaves of (R. prinoides) than in the leaves of most other medicinal plants.

Ethiopian linseed accumulates relatively higher amounts of K and Pb among the essential and nonessential metals, respectively. The non-essential heavy metal, Cd, was not detected. The contents of minerals in Ethiopian linseed are within the daily recommended level and thus advisable as healthy food for treatment of different health complications. C. macrostachyus (an Ethiopian traditional medicinal plant) accumulated appreciable amounts of major and traces metals in its leaves. Ca and Mg were the most abundant nutrients followed by, Fe, Mn and Zn. All the metals enriched in the extract proportionally with extraction time but extent of leaching in the extract was significantly pronounced for the trace metals and the toxic metals (Cd and Pb) for prolonged period of infusion. Thus, beside its medicinal value, C. macrostachyus leaves can be good source of mineral nutrients.

Ethiopian fenugreek seeds are a good source of essential metals. However, they also contained large amounts of the toxic metals Cd and Pb and therefore should not be consumed daily. Prolonged accumulation of heavy metals ingested via foodstuffs may lead to chronic effects on the kidney and liver of humans and cause the disruption of numerous biochemical processes, leading to cardiovascular, nervous, and kidney and bone diseases.

Ethiopian khat varieties contains appropriate concentration of essential major, minor, and trace metals, and they could be source of dietary minerals and trace metals. The toxic heavy metals (Cd and Pb) were found to be too low to be detected by the available technique (FAAS) indicating that the widely chewable and commercially available Ethiopian khat are free from environmental pollution due to non-essential heavy metals. Ethiopian (Cannabissativa L) contains higher levels of essential metals Ca, Zn and Cu. But highly toxic metals like Pb and Cd were also detected in (Cannabissativa L) The content of both toxic metals are beyond permitted limits set by WHO. This will further lead to the harmful effect of (Cannabis sativa L). on human health. In general, the levels of metals in the Ethiopian cannabis are comparable to those reported in the literature from other countries.

The heavy metal component of Ethiopian tobacco is comparable with that of other countries’ tobacco. Pb was not detected in the Ethiopian tobacco leaves which make the Ethiopian tobacco free this toxic heavy metal. Cd concentration in Ethiopian tobacco was found to be comparable with industrialized countries and greater than that naturally available in the soil. The study also revealed that there is large difference between the heavy metal content of raw tobacco leaves and processed tobacco. This indicates that the metal contents that determined in cigarette are not only the content of raw tobacco leaves itself but also the metal originated from contamination of raw tobacco leaves during the process, starting from harvesting to cigarette manufacturing. The study also revealed the dependence of metal accumulation in tobacco leaves on the geographical origin in which tobacco plant grows.

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Application of Radioimmunoassay for Livestock Fertility Management

Introduction

Assessment of hormones in livestock is widely used to achieve the improvement of both production and reproduction. Conventional ways of (genetic) improvement of livestock are based on selection and crossbreeding programme. These programmes have proved to be very effective, but in general they are time consuming and expensive. Furthermore, heritability estimates are generally very low. For these reasons, much of the current research is directed towards the development of alternative methods of selection, which are based on the correlation between biochemical parameters such as hormones, enzymes, metabolites etc and economically important traits. Much of this type of research has initially been directed to the study of blood types and protein polymorphism in which markers are most likely linked to the action of a single gene or a few genes only. These marker genes may be coupled to other genes which are relevant to economically useful traits, and therefore can be used, in principle, as a basis for selection. Some hormones considered to be possible markers for production traits are thyroid hormones, growth hormone, insulin, prolactin, corticosteroids and androgens. The hormones used for markers of reproductive status are LH, testosterone, progesterone, estrogen, estrogen sulphate etc. An important step towards an improved understanding of reproductive physiology was the introduction of highly specific and sensitive radioimmunoassay technique in the early 1960s. Earlier bioassay methods of hormone estimation were not so much reliable and accurate. Radioimmunoassay (RIA) and other related techniques now a day’s allow measurement of hormones and other substances with a sensitivity up to the level of pico gram. Therefore RIA techniques become so popular over the others.

Monitoring of Fertility by Measurement of Hormones: Measurement of progesterone, estrogen, oestrone sulphate, pregnancy specific protein and pregnancy associated glycoprotein has found practical application as a method for improving reproduction in farm animals.

Expression of Estrus

Poor expression of estrus is one of the major factors hampering the efficient utilization of tropical cows and buffaloes. Estrus is traditionally observed by behavioral symptoms which, however is practically very difficult because the overt signs are of low intensity and of short duration. Occurrence of ovulation which is not preceded by overt behavioral estrus symptoms is quite common in tropical animals during stressful summer and non-stressful winter months. Progesterone serves as a marker for determination of functional status of corpus luteum and diagnostic tool for identifying ovarian condition such as estrus confirmation, differentiating types of cysts, silent estrus and lack of cyclicity in cattle and buffaloes. Estrogen is essential for expression of estrus whereas progesterone is required for preparation of uterus for implantation and maintenance of pregnancy. This is because progesterone levels are low around the time of ovulation and high during luteal phase of estrous cycle or during pregnancy and declined to basal level on the day of parturition. Mean plasma progesterone concentration declined from 1.14 ng/ml on day 4 prior to estrus to less than 0.4 ng/ml on the day of estrus and then rose to 2.73, 1.84 ng/ml on day 10 and 8 of the cycle in cattle and buffaloes, respectively [1-3] .

Mean progesterone level was lowest on the day of estrus and rose to a peak level during mid luteal phase of cycle which then declined to a basal level on the day of next estrus in cattle and buffaloes that exhibited overt and silent estrus, respectively [4,5]. The overall plasma progesterone levels in cattle and buffaloes that exhibited silent estrus were lower compared to those in overt estrus and might be responsible for poor expression of estrus. Estradiol levels are maximum on the day of estrus and declines gradually with a minimum concentration on day 12-14 of the cycle. In males, testosterone is responsible for sexual maturity and sperm production. In infertile animals or in case of oligospermic animal, testosterone levels remain low or undetectable. The economic benefit of progesterone based estrus confirmation has not been quantified. Since the proportion of cows not in or near estrus when inseminated varies among herds from 0 to 60% [6], a substancial benefit might be expected, especially on the farms with a high error rate for estrus detection. This benefit should result from reduction in calving interval and the number of inseminations per conception [7] demonstrated a strategy of on farm milk progesterone testing on day 19 after insemination, followed by prostaglandin treatment of non pregnant cows will be profitable, but only if the efficiency of detection of estrus among cows diagnosed non pregnant is increased by more than 20% and if the error rate in pregnancy is less than 3%. To achieve the latter, the cows which are non pregnant on the basis of on farm progesterone test should be checked again for pregnancy by other methods.

Diagnosis of Pregnancy

Early detection of pregnant and non-pregnant livestock has become a key to good breeding management because it is an essential factor for monitoring and controlling fertility. Many new and old technologies are available to identify pregnant and nonpregnant animals early post service. The methods of pregnancy diagnosis are divided into direct and indirect methods. The direct pregnancy diagnosis methods include – transrectal palpation and ultra sonography. Pregnancy diagnosis by rectal palpation have been reported during early pregnancy in cattle [8,9], buffalo [10], sheep [11] and pig [12]. Traditionally, to confirm pregnancy at about day 30 of gestation onwards, the practitioners have relied on the palpation of the amniotic vesicle and slipping of the chorioallantoic membranes between the thumb and forefinger. Palpation technique detects pregnancy with an accuracy of 66 to 100% from Days 49 to 109 of gestation, however it has low accuracy (17 to 57%) for determining multiple fetuses.

Transrectal ultrasonography has been used for pregnancy diagnosis in cattle [13], buffalo [10-14], sheep [8] and goat [14- 17]. Transrectal ultrasonography identifies the embryonic vesicle as early as Day 12 after mating but the sensitivity of this technique for pregnancy is very low (12%) earlier than 25 days after mating. Transrectal ultrasonography for pregnancy diagnosis offers some advantages over palpation per rectum: earlier diagnosis of pregnancy/non-pregnancy, determination of embryo/fetus viability, reduction of misdiagnosis and reduction of “potential” iatrogenic embryo/fetal attrition. The currently available indirect methods of pregnancy diagnosis include measurement of hormones such as progesterone [18-20], estrone sulphate [21,22] and pregnancy specific proteins such as pregnancy-associated glycoproteins [23,24] the early pregnancy factor [25] and interferon-tau [26,27]. Early pregnancy diagnosis will assist dairy producers in managing open cows and improving reproductive performance and economics of their herd.

Progesterone

Measurement of progesterone by RIA has been widely used for verifying whether cows are in estrus at the time of insemination or early diagnosis of pregnancy in cattle, buffaloes, goats and pigs. Progesterone levels elevate during the mid cycle of each reproductive cycle and during the entire gestation period. If the cow is not pregnant, the corpus luteum regresses and progesterone levels decline to low levels about 2 days before the cow comes into heat again. However, if the cow becomes pregnant, the corpus luteum continues to function and progesterone levels remain high throughout gestation. In pregnant cow, progesterone values in peripheral plasma increase with the development of corpus luteum up to conception (5-10 ng/ml) on days 15-20 after conception; these concentrations remain constant until shortly before parturition. Using progesterone assay pregnancy can be predicted between 68 to 95% and this test gives best accuracy after three weeks of pregnancy. Studies in the bovine estrous cycle indicate that the milk or serum progesterone concentrations reach a maximum value 13- 14 days after estrus, and if the animal is pregnant, these continue to remain elevated up to day 21 after fertilization [28] and beyond.

These high levels of progesterone in serum or milk between days 18 and 24 after insemination form the basis of establishment of pregnancy in cattle [29,30]. Since Laing and Heap 1971 first described the use of milk progesterone measurements as an early indicator of the reproductive status of the lactating dairy cow, there has been considerable research on the development of sensitive and reliable assays for progesterone in milk Shemesh et al. [30] proposed that the difference in peripheral plasma progesterone levels between pregnant and non-pregnant cows, 19 days after insemination, can form the basis for a very early pregnancy test. Laing and Heap 1971 first documented this in milk to diagnose cows in early pregnancy. Conception extends the life of the corpus luteum (CL) by preventing the luteolytic mechanism from being triggered, thus prolonging and maintaining its functional characteristics, ensuring continued high progesterone levels [31]. In buffalo cows, the progesterone levels in milk are four to five times higher than those in blood plasma [32,18]. Like cattle, buffaloes too can be accurately diagnosed as non-pregnant by determination of plasma progesterone concentrations 21 days after insemination [33] also reported that progesterone concentration in the milk of pregnant buffaloes was significantly higher than that in non-pregnant animals on Day 20 and the difference between the two increased with time after insemination. The detection of non-pregnant animals was 100% successful at all times but the diagnosis was correct for 66, 68, 81 and 83% of animals tested on Days 20, 24, 28 and 40 respectively and predicted as pregnant.

Shemesh[30] 18-22 day reported that plasma and milk progesterone concentrations in pregnant sheep 18 22 days after mating were similar, about 3.7 ng/ml whereas values in nonpregnant sheep were less than 1 ng/ml. The accuracy was 92- 100% for ewes diagnosed non-pregnant in the breeding season, but for ewes tested in the non-breeding season the diagnosis of non-pregnancy according to milk progesterone levels was only 50% accurate. Progesterone concentrations decrease sharply during regression of the corpus luteum (CL) in the non pregnant doe (one or two days prior to estrus) and return to higher levels within four days following estrus [34-36]. The pregnant doe has high progesterone concentrations during the same time period since the CL does not regress. Recently, Mondal, et al. [37] observed that plasma progesterone concentration declined from day 25 pre partum abruptly to the day of kidding and remained at basal level up to day 25 postpartum in Black Bengal goats.

Estrone Sulphate

Estrone sulphate is a conjugated estrogen that can be detected in maternal plasma, serum, milk and urine. It is produced by the feto maternal axis or the conceptus and its presence is indicator of pregnancy. Actually the viability of fetus is determined by measurement of estrone sulphate. It is detected at day 72 in plasma and between 105-112 days of gestation in milk of cows. Estrone sulphate test can reliably be used to diagnose the pregnancy beyond day 100 of gestation. This test can also be used to diagnose both singlet and multiple calf pregnancies after 110 days of pregnancies. Estrone sulphate test can reliably be used to diagnose the pregnancy beyond day 100 of gestation. This test can also be used to diagnose both singlet and multiple calf pregnancies after 110 days of pregnancies. Estrone sulphate in milk of cows rises from 30 pg/ml to 151 pg/ml in whey between days 41 and 60 of gestation to reach a maximum concentration of about 1000 pg/ml at days 220-240 of pregnancy [37]. Examination of the ranges of ES concentrations in milk sampled from non-pregnant and pregnant cows indicated that all non-pregnant cows and 46% of cows <120 days pregnant had milk ES concentrations <125 pg/ml. However, only 4% of cows ≥ 120 days pregnant had milk ES concentrations <125 pg/ml. The levels of estrone sulphate in different maternal body fluids, namely, milk and blood plasma, can be utilized as the criteria for confirming pregnancy by after 110 day insemination in bovine species [38].

In Murrah buffaloes the levels of of estrone sulphate were below detection levels (<50 pg/mL) during the first two months, followed by sharp increase in the fourth month and values stabilized after reaching the highest levels in the sixth month of pregnancy [39]. Hung and Prakash recorded a progressive increase in estrone sulfate concentrations in buffalo plasma after the 4^th or 5^th month of pregnancy. In an another study by Tsang [40], estrone sulphate was detectable around Day 70 of gestation with value ranging between 0.1 to 0.7 ng/ml, then its level increased steadily till 2 days before parturition when an upsurge was seen (15-50 ng/ml). On Day 85 of gestation, there was a significant difference in the level of estrone sulphate between pregnant and non-pregnant ewes. In goat, oestrone sulphate concentrations began to rise from about Day 40 and reached a plateau of about 19nmol/l by Day 120 [41]. This value was maintained until about 20 days pre partum when there was a slight decline in concentration, rising by 2-fold about 24 h before parturition. The accuracy for detection of non-pregnancy was only 44 % whilst for detection of pregnancy it was 87.9% using the cut-off value of 0.1ng/ml [42]. Estrone sulphate based on farm pregnancy diagnosis is also possible in foecal sample of pigs. The concentration of total unconjugated oestrogens and estrone are consistently higher between days 24 and 30 in pregnant sows compared to non pregnant sows [43].

Pregnancy Associated Glycoprotein (PAG)

Pregnancy-associated glycoproteins (PAG) belong to a large family of inactive aspartic proteinases expressed by the placenta of domestic ruminants including cows, ewes, and goats [44]. PAG are synthesized by the mono- and binucleate trophoblastic cells of placenta and some of them are secreted in maternal blood from the moment when the conceptus becomes more closely attached to the uterine wall and formation of placentomes begins [45]. Among these glycoproteins, Butler, et al. [46] detected two pregnancy-specific proteins in the sera of pregnant cows – 65-70kDa and a 47-53kDa protein. Of these, the former showed an immune reaction similar to that of a 1-fetoprotein, while the latter showed no reactivity with known proteins and it was given the name “protein B” or the “pregnancy-specific protein B” (PSPB) in bovines. Pregnancyspecific protein-B (PSPB) was the first pregnancy-specific marker identified in cattle [46] and was later found to have the same N-terminal amino acid sequence as bovine PAG-1. The PAG family were isolated from cotyledons of cow [47-49], ewe [47,50], goat [51] and buffalo [52].

Development of specific RIA and EIA for the presence of pregnancy associated proteins of feto-placental origin in the maternal serum 3-4 wks after conception has been used as a serological marker for pregnancy diagnosis in cattle [53], sheep and goats [54]. During gestation of cow, the concentrations of PAG are detectable as early as from the 19th to 22nd days after the conception, to reach concentrations from 3 to 6 ng/ml in the neighborhoods of the 33rd to 37th days of gestation [55,56] reported that PAG concentrations increased continuously from day 20 of pregnancy until day 240 followed by a dramatic increase in the last ten days of pregnancy with maximum concentrations between day 5 and day 1 prepartum. In the bovine species, two different patterns of expression were found – those PAG that are expressed predominantly in binucleated cells (PAG 1-subgroup) are invariably absent in term placenta, whereas those PAG expressed more uniformly throughout trophoectoderm (PAG 2-subgroup) are detectable at all stages of pregnancy [57]. For example, boPAG-9 is expressed predominantly in early pregnancy, being detectable at Day 25 and declining as pregnancy progresses until being undetectable at term. On the contrary, boPAG-1 is not detectable at Day 25 but becomes prevalent at later stages, although it is absent in term placenta. Bovine PAG-2, -8, -10, and -11 are detectable throughout gestation and they are the only PAG present in term placenta. PAG measurement has also been used in several studies to monitor pregnancy failure during the late embryo and early foetal period [53,58].

In sheep the plasma PAG profiles are characterized by an initial increase between the 3rd and 4th week, followed by further gradual rise up to the 9th week of pregnancy [59]. Between the 9th and the 19^th week the level remained constant, thereafter a drastic surge occurs, reaching a peak at parturition. Recently Rovani et al. [54] detected pregnancy accurately using bovine ELISA kit in sheep 33 days following mating, while PAGs levels from the previous gestation are no longer detected from 21 days postpartum. The accuracy of the ELISA test was 96.1% from 33 days of pregnancy until lambing. In goat’s perusal of literature revealed a different profile of PAG with a significant first increase between day 21 and day 28 and maximum levels between the 5^th and 8^th week of pregnancy. Thereafter, PAG levels decreased slowly until parturition [17] reaching basal levels in the 4^th week postpartum. Recently [57] was able to diagnose pregnancy in Boer goats from day 28 of pregnancy onwards using an ELISA based on antibodies raised against caprine or ovine PAG. With the antibody rose against caprine PAG a steep increase to a peak level of 69±9ng/ml on day 56 of pregnancy was followed by a gradual decline to 16±3 ng/ml at parturition and 0.3±0.07ng/ml four weeks postpartum. With antibovine PAG, the PAG level increased to a maximum of 3.1±0.2 ng/ ml on day 105 of pregnancy and fluctuated around 3ng/ml until the end of pregnancy suggesting its ability to diagnose early pregnancy and its possible use to evaluate feto-placental well-being [60,61].

Interferon-Tau

In ruminants, the anti luteolytic hormone for pregnancy recognition and maintenance of functional corpora lutea (CL) is interferon tau (IFN-τ) [62,63]. It is secreted in large quantities by the mononucleate cells of the trophectoderm as the blastocyst begins to elongate at about days 13 in cattle with a peak production when the concepts reaches its maximal size [64]. The secretion of IFN-τ by mononuclear cells of the ovine trophectoderm is developmentally regulated with onset of secretion occurring as large spherical blastocysts transition to tubular and elongated filamentous forms between days 10 and 21 of pregnancy. In sheep, it is secreted between days 10-21 by the mononuclear trophoblast cells. On days 11-16, the PGF_2α concentrations are the same in pregnant and non-pregnant animals, but pregnant animals administered PGF2α on day 19 or 20 do not return to estrus. It is the pregnancy recognition hormone in sheep and other ruminant that acts to silence the transcription of estrogen receptor alpha (ESR1) and, therefore, ESR1-dependent expression of the oxytocin receptor (OXTR) gene in both uterine LE and superficial glandular epithelium (sGE), hereafter referred to as LE/sGE.

This abrogates development of the endometrial luteolytic mechanism that requires oxytocin-induced release of luteolytic pulses of prostaglandin F2α (PGF) by uterine LE/sGE; however, circulating concentrations of PGF are greater in pregnant than cyclic ewes due to continued expression of prostaglandin endoperoxide synthase 2 (PTGS2). IFN-τ is a member of the Type I IFN family that acts differentially on the endometrial luminal epithelium (LE), glandular epithelium (GE) and stroma to regulate expression of a number of IFN-stimulated genes (ISGs) that are hypothesized to play roles in the endometrial differentiation and conceptus implantation [65-68]. It has been demonstrated that bIFN-τ has several effects on the endometrium that result in decreased PGF2α secretion in the pregnant cow and maintenance of the CL. For example, IFN-τ reduces estradiol receptor number and thus prevents an estrogeninduced increase in oxytocin receptor number [69-74]. Moreover, rbIFN-τ inhibits the oxytocin- and/or phorbol ester-induced increase in phospholipase A2, cyclooxygenase-2 and prostaglandin F synthase expression in bovine endometrial cells.

Conclusion

Exploration of fundamental endocrine involvement in reproductive processes is essential for optimum reproductive management strategies and paradigm to overcome the infertility in domestic ruminants. Reliable techniques for early detection of pregnancy aid in culling or rebreeding of animals and provide a valuable tool for controlled breeding programs. Traditional methods of visual observation, abdominal palpation, service records and non-return to estrus are not reliable means of diagnosing early pregnancy. The review has lime lighted the use of hormonal assays for detection of estrus and early pregnancy in cattle, buffalo, sheep, goat and pig with their accuracy. It is concluded that micro quantitation of hormones has come as a revolutionary breakthrough in understanding animal reproduction function and is an important research tool for augmenting both production and reproduction of domestic animals.

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Fluoride Content in Ground Water¬ in Mahendergarh (Haryana)

Introduction

Water, the precious gift to nature of human being is going to be polluted day by day with in increasing urbanization. Although three fourth part of earth is being surrounded by water but a little portion of it can be used for drinking purpose. In India around 62.5million people are suffering from disorder of teeth or bones through fluorosis [1]. Seventeen states in India have been identified as endemic for fluorosis & Haryana is one of them. Although fluoride enters through food, water, industrial exposure, drugs, cosmetics etc. Drinking water is the major contributor (70-90% of daily intake) [2]. Due to its strong electro negativity; fluoride is attracted by positively charged calcium in teeth and bones. The major1health problem is caused by fluoride are dental fluorosis, teeth mottling, Skelton fluorosis and deformation of bones in children as well as adults [3]. According to WHO [4], permissible limits for fluoride in drinking water is 1.0mg /l [4] where as USPHS [5]. The maximum allowable concentration for fluoride in drinking water in Indian condition comes to 1.0 mg/l while as per Indian standards it is 1.5mg/l [6-8]. The routine monitoring of water can assure the populace that the quality of their drinking water is adequate. It can also be beneficial in detecting deterioration in the quality of drinking water and facilitate appropriate timely corrective actions with minimal negative impacts on population health.

Materials and Methods

Water Sampling: A total of 10 ground water samples taken from ten locations of Mahendergarh district were collected in polythene bottles which were cleaned with acid water, followed by rinsing twice with distilled water. The water samples are chemically analyzed. The analysis of water was done using procedure of standard methods.

Study Area: Mahendergarh district, which is one of the 21 districts of Haryana state in northern India. The district occupies an area of1, 859 km2.The district has a population of 812,022(2001 census). Narnaul Town is the administrative headquarters of the district. Mahendergarh is one of the very few districts in India where the name of the district and its town are different. As of 2011 it is the third least populous district of Haryana (out of 21), after panchkula and Rewari. The district lies between north Study Area: Mahendergarh district, which is one of the 21 districts of Haryana state in northern India. The district occupies an area of1, 859 km2.The district has a population of 812,022(2001 census). Narnaul Town is the administrative headquarters of the district. Mahendergarh is one of the very few districts in India where the name of the district and its town are different. As of 2011 it is the third least populous district of Haryana (out of 21), after panchkula and Rewari. The district lies between north latitude 270 to 280 26 and east longitude 750 56’ to 760’51’.

Methodology: F-Spectophotometrically using ELICO-52 UV Spectrophotometer [9].

Results

Most of the water sample, collected from the different location in Mahendergarh do not meets the water quality standards and many other quality parameters. Hence it is not suitable for consumption with any prior treatment.

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A Research Note on the Treatment of Naturally Induced Gangrenous Dermatitis in Chickens by Copper Sulfate |

Opinion

The disease condition known as gangrenous dermatitis, avian malignant edema, necrotic dermatitis, gangrenous cellulites or gangrenous dermatomyositis has been described by many investigators [1,2]. Gangrenous dermatitis is a disease of young growing chickens characterized by gangrenous areas of the skin and by severe underlying infectious cellulites. Outbreaks often occur in excessively warm, humid houses [3]. Losses in affected flocks are ranging between1 and 60% [4]. Clostridium perfringens type A and Clostridium septicum were blamed as the main etiological agents for this infection [4]. Staphylococci spp and E. coli are also incriminated in this infection [1-3].

Beemer [5] reported on an epizootic of dermatitis in chickens caused by the yeast Rhodotula mucilaginosa. Bal and Dabiya [6] reported on Aspergillus fumigates as the etiological agent of avian dermatitis. The condition occurs in broiler chickens and is frequently associated with high encounter outbreaks in replacement laying stock from 4-16 weeks of age [7] who also concluded that drinking water medication with antibiotics in view of complicated etiology is disappointing. The present note dealt with the use of copper sulfate in treating gangrenous dermatitis in chickens. In the present investigation 3 separate replacement laying chicken flocks suffered from total losses of 2.6. 19.2 and 4.5% in the 3 flocks within a period of 6, 10 and 12 weeks respectively due to gangrenous dermatitis did not respond to treatment with zinc bcitracin, ampicillin, oxytetraccline or streptomycin.

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A Case Study: Polychlorinated Dibenzo-p-dioxins, Polyclorinated Dibenzofurans, Polychlorinated biphenyls, and Polybrominated diphenyl ethers in the bone marrow and lymph nodes of adult reindeer (Rangifer tarandustarandus L.) hind in Finland |

Introduction

It is typical for POPs that they are transported long distances from source to sink areas. Reindeer (Rangifer tarandustarandus L.) is considered to be one of the most important indicator species for environmental toxic contaminants because it uses natural pastures and is exposing continuously to environmental contaminant deposition [1]. The immune system is one of the most sensitive targets for the toxic effects of TCDD and structurally similar POPs [2]. The Aryl Hydrocarbon Receptor (AhR) is important factor in immunotoxic mechanism of POPs. Many pathways of cellular activation, proliferation, and survival have the potential to be influenced by AhR activation, leading to alterations in the function of the immune system [3,4]. We report PCDD/Fs, PCBs, and PBDEs content of tissues relating to immune system and hematopoiesis, lymph nodes and bone marrow, of adult reindeer hind. The congener specific profiles are explored and compared between these two tissue types.

Material and Methods

An adult reindeer hind (age > 10 years) from Finnish reindeer management area (southern site) was slaughtered and lymph nodes and bone marrow samples were immediately extracted for analysis. High resolution gas chromatography – high resolution mass spectrometry (HRGC/HRMS) was used to analyses of POPs. Analyzes were performed at the National Institute for Health and Welfare (THL), in the Unit of Chemical Exposure, in Finland. The Unit is a FINAS accredited testing laboratory (No T077) according to EN ISO/IEC 17025 requirements.

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