Several nucleic acid-based assays have been developed for detecting Anaplasma marginale and Anaplasma centrale in vectors and hosts, making the choice of method to use in endemic areas difficult. We evaluated the ability of the reverse line blot (RLB) hybridisation assay, two nested polymerase chain reaction (nPCR) assays and a duplex real-time quantitative polymerase chain reaction (qPCR) assay to detect A. marginale and A. centrale infections in cattle (n = 66) in South Africa. The lowest detection limits for A. marginale plasmid DNA were 2500 copies by the RLB assay, 250 copies by the nPCR and qPCR assays and 2500, 250 and 25 copies of A. centrale plasmid DNA by the RLB, nPCR and qPCR assays respectively. The qPCR assay detected more A. marginale- and A. centrale-positive samples than the other assays, either as single or mixed infections. Although the results of the qPCR and nPCR tests were in agreement for the majority (38) of A. marginale-positive samples, 13 samples tested negative for A. marginale using nPCR but positive using qPCR. To explain this discrepancy, the target sequence region of the nPCR assay was evaluated by cloning and sequencing the msp1β gene from selected field samples. The results indicated sequence variation in the internal forward primer (AM100) area amongst the South African A. marginale msp1β sequences, resulting in false negatives. We propose the use of the duplex qPCR assay in future studies as it is more sensitive and offers the benefits of quantification and multiplex detection of both Anaplasma spp.

Objective: The identification of meat species in meat products is important for protection of human health, economic reasons, religious factors and for controlling the compliance with food regulations. For this purpose, DNA must be obtained in good quality and quantity. The aim of this study was to compare different DNA isolation methods from different meat products.Materials and methods: Comparison among different DNA isolation methods was done. DNA was isolated from different meat products (e.g., sucuk, salami, sausage, braised meet, meatball and pastrami). The methods included phenol/chloroform, DNA isolation kit, Cetyl Trimethyl Ammonium Bromide (CTAB) and Dodecyle Trimethyl Ammonium Bromide (DTAB).Results: Although DNA was obtained from all of these methods, the phenol/chloroform and DNA isolation kit methods were found to be the most effective methods for obtaining high quantity DNA. RNA contamination was determined to be common in DTAB method. High quantity of DNA and RNA contamination in terms of quality was detected in CTAB method. Ruminant specific 16S rRNA primer was used to amplify genomic DNA by polymerase chain reaction and all samples were amplified except for some samples of DTAB.Conclusion: DNA isolation kit, another best method, is recommended due to quality and quantity of DNA for researchers who do not want that phenol/chloroform method have toxic substances. This study is also the first study in which DTAB method is used for DNA extraction from meat products.http://doi.org/10.5455/javar.2016.c175

Objective: Brucellosis is a highly contagious zoonosis with global distribution. The disease remains endemic in many countries including Iran, while its seroprevalence in endemic area is not well documented. We aimed to determine the seroprevalence of brucellosis in sheep and goats in Hamedan province, west of Iran.Material and methods: A total of 3,250 blood samples from 2,550 sheep and 700 goats were collected randomly. All samples were analyzed for the presence of Brucella antibodies using Rose Bengal, Wright standard tube agglutination and 2-mercaptoethanol agglutination tests.Results: The seroprevalence rate of brucellosis in animals and flock level were found in 4.6% and 13.6% of goats and 3% and 27.9% of sheep, respectively. No evidence of correlation between gender and Brucella infection rate were found in animals (P>0.05). Statistical significant differences was seen between age groups and infection rate in goats (P=0.033, OR=2.1); unlike to sheep (P=0.373). Also, the infection rate in nomads population of sheep was higher than fix location animals (P=0.003; OR=1.9); unlike to goats (P=0.195). In animals with history of abortion and vaccination against brucellosis, seroprevalence rate was significantly lower than other (P<0.05).Conclusion: This is the first report of brucellosis in sheep and goats in Hamedan province. The design of a comprehensive control program including vaccination, screening, and culling of brucellosis-positive animals is recommended.http://doi.org/10.5455/javar.2016.c179

Objective: This study was carried out to investigate the effects of estradiol benzoate (EB) injection to male rabbits on glucose, total-protein, albumin, calcium concentrations and prostate tissue, and the role of prolactin as an important mediator of estrogen action in prostate.Materials and methods: Fifty four adult male rabbits were used in this study. The rabbits were randomly divided into two groups. Group A contained 36 male rabbits, which were further randomly divided into four sub-groups, three of them contained 10 rabbits and one sub-group contained 6 rabbits as control. Group B contained 18 male rabbits, which were divided randomly into three equal sub-groups. Three sub-groups of Groups A and B were treated once each on alternative day with the intramuscular injections of EB dosed at 40, 80 and 120 µg/rabbit, respectively for 20 days, whereas the fourth sub-group of Group A received no estradiol, and Group B received 1 mg Bromocriptine Mesilate in addition to EB through oral route on each alternative day. Blood samples were collected for measuring glucose, t-protein, albumin and calcium levels. Prostate tissue samples were collected from all the rabbits for histological studies.Results: Glucose was significantly (P?0.05) increased as a result of 80 µg EB injection, while significantly (P?0.05) decreased due to 40 and 120 µg EB injection. Total protein significantly (P?0.05) increased due to injection of 40 µg EB, however t-protein was not changed due to 40 and 120 µg injection. On the other hand, the results of albumin and calcium were not affected (P?0.05) by EB. In prostate tissues, EB induced hyperplasia with dysplasia or dysplasia only, but this effect was mild due to inhibition of prolactin.Conclusion:  The injection of EB to male rabbits increased or decreased glucose level, increased t-protein level mildly or not changed, while albumin and calcium levels were not affected. EB induced hyperplasia on prostate tissue, and this effect was reduced by prolactin inhibition indicating that prolactin might have a role on the action of estrogen.http://doi.org/10.5455/javar.2017.d184

Objective: This study envisages dynamics and factors affecting fecal oocyst counts (FOC) in natural infection in goats in semi-arid India.Materials and methods: A total of 1152 fecal samples from Jamunapari goats in semi-arid India were collected, processed and examined for fecal oocysts over a period of 3 years for prevalence and severity of the disease through FOC using modified Mc Master Technique. The log transformed FOC data on 912 animals from 59 sires were used for subsequent analyses for genetic and non-genetic factors affecting FOC. Fixed effects included were years of collection (1-3), seasons of collection (summer, rainy, winter), sex (male, female), age group (0-3, >3-6, >6-12, adults). Least squares analysis of variance for fitting constant was applied to data set.Results: The overall prevalence of coccidian infection in goats was 86.71%(n=999/1162). Highest incident was found in winter season (91.74%) and >6-12 M age (97.95%). Eimeria arloingi and E. ninakohlyakimovae were the most frequently occurring species. The heritability of FOC was found to be 0.06±0.06. The effect of sire on FOC was not significant; however, effects of animal age, year, season of collection and sex on FOC were found to be significant.Conclusion: Coccidiosis in goats is seasonally occurring disease, most commonly affecting animals of less than one year age. FOC in coccidiosis affected animals is lowly inherited trait for selection of goat against this disease.http://doi.org/10.5455/javar.2017.d190

Objective: This research was designed to establish the system of isolation and culture of Anas platyrhynchos (duck) amniotic fluid stem cells (DAFSCs), and to explore its biological characteristics and differentiation ability in vitro.Material and methods: Main experimental reagents contained L-DMEM, fetal bovine serum, chicken serum, EGF, bFGF, L-glutamine, trypsin, rabbit anti-chicken CD44, CD73, CD105, nanog and SSEA-4 (Abcam, USA), FITC conjugated goat anti-rabbit secondary antibody IgG, DAPI, Trizol, inverse transcription kit, Propidium iodide, IBMX, INS, dexamethasone and indometacin. Cultivation system included L-DMEM with 10% FBS, 5% chicken serum, EGF 10 ng/mL, bFGF 10 ng/mL and 1% L-glutamine, and was cultured under 37°C, 5% CO2 and saturated humidity. Immunofluorescent detection is used to detect cell surface markers, while RT-PCR was used to detect related gene expression. Cell cycle was detected with Flow Cytometer and was analyzed by ModFitLT 2.0, induced differentiation, and Oil Red O staining.Results: More DAFSCs were gained via super-centrifugation and thermoelectric methods cost effectively. DAFSCs could go down to the future generation at passage 23(P23). CD44, CD73, CD105 and SSEA-4 were detected as positive with immunofluorescence histochemistry. GAPDH, GDNF, rex1 and JAG1 were detected as positive with RT-PCR. Cell cycle was detected on flow cytometer. Tentative exploration of differentiation ability that DAFSCs could be induced into adipocyte in vitro.Conclusion: DAFSCs can be isolated from matrix that have strong self-renewal capacity in vitro. DAFSCs can be induced into adipocyte in vitro. These testify that DAFSCs can be an ideal seeded cells having potentials for preservation and utilization of rare genetic resources.http://doi.org/10.5455/javar.2017.d200   XML     PubReader

Objective: The objective of this study was to isolate and identify Pasteurella spp. associated with pneumonic lungs showing respiratory signs of goats in Ethiopia.Materials and methods: A total of 2400 goats that were slaughtered at the Hashim’s Ethiopian Livestock and Meat Export abattoir, Ethiopia were randomly selected for this cross-sectional study during the period of October 2013 to July 2014. Detail ante-mortem, and post-mortem (PM) lesions were inspected, and the suspected samples were collected aseptically from the lungs. Among 2400 goats, 31(1.29%) goats were not slaughtered because these goats showed severe clinical signs. Thus, 2369 goats were slaughtered finally. The collected samples were subjected for isolation and identification of bacterial species following conventional methods such as culture and biochemical examinations.Results: Out of 2400 goats examined, 960(40%) animals showed different abnormal respiratory signs. Based on PM findings, 16.21% (n=384/2369) lungs were found as pneumonic, of which 78.38% (n=301/384) were found to be associated with Pasteurella organism. The overall prevalence of Pasteurella organism (Mannheimia haemolytica and Pasteurella multocida) was 12.71% (n=301/2369). In this study, youngers and goats with medium body condition score (BCS) had greater probability (P<0.05) to be infected by the bacteria though there was no difference in exposure to the organism among goats from Arsi, Bale and Hararghe. On the other hand, out of 301 positive cases, 274(91.03%) were caused by M. haemolytica, and 27(8.97%) were caused by P. multocida isolates.Conclusion: Pasteurella organism especially M. hemolytica is one of the most common causes of pneumonic pasteurellosis in caprine at the study area. So, chemoprophylaxis needs to be given to small ruminants prior to transportation or other stress conditions.http://doi.org/10.5455/javar.2017.d202XML    PubReader

Objective: In this study, a serological survey was conducted in unvaccinated sheep and goat populations at Pyawbwe and Meikhtila townships of Mandalay region in Myanmar to determine the seroprevalence and associated risk factors of foot and mouth disease (FMD).Materials and methods: A total of 110 sheep and 107 goat sera samples were randomly collected from Pyawbwe. Similarly, 108 sheep and 109 goat sera were collected from Meikhtila. All samples were tested for the presence of non-structural protein (NSP) specific antibodies to FMD virus (FMDV) by Ceditest FMDV-NSP Enzyme-lined Immunosorbent Assay (ELISA), and were confirmed by Liquid Phase Blocking ELISA (LPB ELISA) .Results: Overall seroprevalence was 42.4%(n=184/434) by Ceditest-NSP ELISA, and 46.8%(n=203/434) by LPB ELISA against FMDV serotype O. The presence of antibodies against FMDV serotype O was higher (P<0.01) as compared to those of serotype A and Asia-1. The seroprevalence in Meikhtila (49.77%) was higher (P<0.01) than that of Pyawbwe (35.2%). The seropositivity in sheep and goats that were in-contact (77.19%) with infected cattle and pigs was higher (P<0.01) as compared to those in-contact with non-infected animals (37.14%). Similarly, the seropositivity in sheep and goats from high animal trade areas (49.4%) was higher (P<0.05) than that of those from low animal trade areas (37.97%).Conclusion: Rearing of sheep and goats in-contact with FMDV-infected cattle and pigs, and high animal trading areas are the major associated risk factors for FMDV infection for sheep and goats in the study areas in Myanmar.http://doi.org/10.5455/javar.2017.d204XML   PubReader

Objecitve: Maggot wound is common in domestic and pet animals but report on maggot wound treatment in wildlife species is scanty. The study reported here the surgical and conservative management of maggot wounds including anesthetic protocol and postoperative care in two Bengal tigers (Panthera tigris tigris).Materials and methods: One female and one male tiger were presented with maggot wounds for treatment at the Bangabandhu Sheikh Mujib Safari Park, Gazipur, Bangladesh. Tigers were anesthetized with combined injection of xylazine (dosed at 1.0 mg/kg bwt, IM) and ketamine hydrochloride (dosed at 3.5 mg/kg bwt, IM). Superficial maggots were removed from wounds using sterile tissue forceps. Gauze soaked in oil of turpentine was allowed to remain in each wound pocket for 5 min for the removal of deep-seated maggots. Finally, wounds were dressed with tincture iodine to clean out the dead tissue debris and to induce inflammation for rapid healing. A single subcutaneous injection of ivermectin (dosed at 200 µg/kg bwt, IM) was given in each tiger. In addition, long acting oxytetracycline (dosed at 10 mg/kg bwt, IM) on 48 h interval for six days, chlorpheniramine maleate (dosed at 1 mg/kg bwt, IM) once daily for three days, and ascorbic acid (dosed at 250 mg/tiger, IM) once daily for seven days were administered in both tigers.Results: The tigers were recovered successfully without any complications in two weeks following treatment.Conclusion: Surgical management using oil of turpentine and tincture iodine along with parenteral administration of ivermectin, long acting oxytetracycline and chlorpheniramine maleate are effective for successful management of maggot wounds in Bengal tigers.http://doi.org/10.5455/javar.2017.d187

Objective: Molecular based study was conducted to determine the associated serotypes in the reemergence of Foot and Mouth Disease (FMD) outbreak in Assiut governorate, Egypt during 2014 and 2015.Materials and methods: One hundred and twenty cattle with clinical signs suggesting their infection by FMDV were examined clinically and twenty three of them were used for confirmation by laboratory diagnosis. Different clinical samples including vesicular fluid and tongue epitheliums were collected and after RNA extraction using commercial kit, RT-PCR was done using different primer sets.Results: Serotype O was detected in 8 samples, 2 of them were also positive for SAT2 serotype. The determination of specific serotype was failed in case of the rest 13 samples although they were positive when tested by the universal primer specific for FMDV. Conclusion:   Serotypes O and SAT2 were the more prevalent serotypes in the current outbreak in Assiut governorate, Egypt.http://doi.org/10.5455/javar.2017.d186