Comparative efficacy of standard AGID and precipitinogen inhibition test with monoclonal antibodies based competitive ELISA for the serology of <Emphasis Type="Italic">Peste des Petits Ruminants</Emphasis> in sheep and goats

This project was conducted to investigate the comparative efficiency of competitive ELISA (cELISA), standard Agar Gel Immunodiffusion Test (AGID) and Precipitinogen Inhibition Test (PIT) for the diagnosis of Peste des Petits Ruminants (PPR) in Pakistan. To deal with this, serum samples from 198 sheep and 82 goats were collected from three different government livestock farms and all the samples were run simultaneously with the three serological tests. The samples found positive for PPR antibodies through cELISA, AGID and PIT were 96 (34.2%), 60 (21.4%) and 72 (25.7%), respectively. Kappa statistics were applied to evaluate the concordance between the laboratory-based test (cELISA) and field-based tests (AGID and PIT). Kappa statistics scores for cELISA versus AGID and PIT were 0.6343 (95% Confidence Interval CI 0.5231–0.7456) and 0.7134 (95% Confidence Interval CI 0.5987–0.8281), respectively, which indicate a “substantial” agreement between cELISA and AGID and “significant” agreement between cELISA and PIT. AGID and PIT revealed relative diagnostic sensitivities with cELISA of 59.3% and 69.7% and relative diagnostic specificities of 98.3% and 97.2%, respectively. The data suggested that for mass screening and control of PPR, these serological tests proved practical in the absence of cELISA since they have high relative diagnostic specificities and a satisfactory relative diagnostic sensitivities.     

Evaluation of foot and mouth vaccination for yak (<Emphasis Type="Italic">Bos grunniens</Emphasis>) in Pakistan

In northern Pakistan, many farming communities rely on domestic yak (Bos grunniens) as a principle source of income. A 2006 participatory disease surveillance report from this region indicated that foot-and-mouth disease (FMD) is the most prevalent annual disease of yak. Our objectives of this study were to determine exposure levels of yak to FMD virus; implement a vaccination program based on current, regional FMD virus serotypes and subtypes; and quantify immune responses following vaccination. Blood samples were used to determine pre-vaccination exposure of animals to FMD virus by antibody presence to non-structural proteins of FMD virus using a 3-ABC trapping indirect ELISA. Vaccine used consisted of FMD serotypes ‘O’ (PanAsia-2), ‘A’ (Iran-05), and ‘Asia-1’ (Shamir), but changed later during the study to match newly circulating viruses in the country (‘O’-PanAsia-2; ‘A’-Turk-06 and Asia-1-Sindh-08). Three hundred sixty-three blood samples were tested from selected villages to determine pre-vaccination FMD virus exposure in yak with an average of 37.7%. Immune responses from initial vaccination and booster dose 30 days later showed clear protective levels (as mean percent inhibition) of antibodies against structural proteins of serotypes ‘O,’ ‘A,’ and ‘Asia-1.’ These responses remained above threshold positive level even at day 210 following initial vaccination. Results of sero-surveillance and anecdotal information of repeated FMD outbreaks demonstrate the persistence of FMD virus of yak in northern Pakistan. Laboratory results and field observations clearly indicated that yak can be protected against FMD with a good quality vaccine with FMD serotype(s) matching current, regionally circulating FMD virus.     

Modulatory function of NMDA glutamate receptor on MC<Subscript>3</Subscript>/MC<Subscript>4</Subscript> receptors agonist-induced hypophagia in neonatal meat-type chicken

Melanocortin 3 and 4 receptors (MC₃R and MC₄R) are known as the main receptors for melanocortin-induced hypophagia in mammalian and poultry. Also, central glutamatergic system has mediatory role on function of the melanocortin system in some brain areas. So, the aim of the current study was to determine the role of MC₃/MC₄ receptors agonist on food intake and its interaction with glutamatergic in 3-h food-deprived (FD₃) neonatal broilers. In experiment 1, chickens were intracerebroventricular (ICV) injected with control solution, MTII (MC₃/MC₄ receptors agonist; 2.45, 4.8 and 9.8 pmol). In experiment 2, control solution, SHU9119 (MC₃/MC₄ receptors antagonist; 0.5, 1 and 2 nmol) were ICV injected. In experiment 3, birds ICV injected with control solution, SHU9119 (0.5 nmol), MTII (9.8 pmol) and co-injection of the SHU9119 + MTII. Experiments 4–8 were similar to experiment 3, except birds injected with MK-801 (NMDA glutamate receptors antagonist, 15 nmol), CNQX (AMPA glutamate receptors antagonist; 390 nmol), AIDA (mGLUR₁ glutamate receptors antagonist; 2 nmol), LY341495 (mGLUR₂ glutamate receptors antagonist; 150 nmol) and UBP1112 (mGLUR₃ glutamate receptors antagonist; 2 nmol) instead of SHU9119. Then, cumulative food intake was recorded until 120 min after injection. According to the results, dose dependent hypophagia observed after ICV injection of the MTII (p < 0.05). ICV injection of SHU9119 significantly increased food intake in birds (p < 0.05). Co-injection of SHU9119 + MTII significantly inhibited MTII- induced hypophagia in neonatal chicks (p < 0.05). In addition, hypophagia- induced by MTII was significantly attenuated with co-injection of MTII + MK-801(p < 0.05). These results suggested MC₃ and MC₄ receptors have inhibitory role on food intake and this effect is probably mediated by NMDA glutamate receptors in neonatal chickens.     

In vitro control of parasitic nematodes of small ruminants using some plant species containing flavonoids

This study determined in vitro anthelmintic efficacy of three plant species: Trema orientalis, Urtica dioica and Zanthozylum capense on nematode larvae of small ruminants. Dried leaf samples (40 g) were extracted in 70% ethanol, in portions of 10 g and concentrated to 100 ml. Half and one quarter of the original crude extract were both made to 100 ml. Rectal faecal material from 10 Merino sheep and 25 Nguni goats was pooled within species and thoroughly hand-mixed. Dung samples, each of 5 g were cultured for 12 days at 27 °C. On day 13, 4 plates were watered and 4 others treated with ethanol to correct for solvent effect on mortality. The design was 2 (animal species)?×?3 (plant species)?×?3 (extract concentrations). In each of three runs, three plates were treated with each crude extract in three incremental concentrations. Surviving L3 larvae were isolated, counted and mortalities became indices of anthelmintic efficacy. Data from nematode larval mortality were analysed to determine the effect of animal species, plant species, concentration and their interactions. Efficacy was affected by concentration (P?=?0.0001), animal species (P?=?0.0046), plant species (P?=?0.0572), the interactions of animal species and concentration (P?=?0.0010), plant species and concentration (P?=?0.0123) and concentration × animal × plant species (P?=?0.0435).     

Equine salmonellosis in southern Brazil

The Salmonella sp. genus is identified in several species, and the zoonosis it causes is one of the most important types worldwide. The specifics of salmonellosis vary according to the function of the serovar involved, the species affected, age and predisposing factors. However, few cases of equine salmonellosis have been reported. This study presents ten confirmed salmonellosis cases in equines in southern Brazil. Six were adult animals with stress factors preceding the disease, while four were foals, three of which presented with hyperacute manifestations. The main clinical signs were diarrhea, anorexia, and hyperthermia. Lesions varied in distribution and severity, although fibrinonecrotic or necrohemorrhagic enteritis was observed in all animals, mainly in the large intestine (large colon and cecum—8/10) and small intestine (3/10). Substantial liquid content, mainly hemorrhagic, was observed in all animals. The most characteristic microscopic lesion was mucosa necrosis, which is often accompanied by fibrin deposition, followed by necrosis of follicular centers and vascular changes. Bacterial isolation revealed seven isolates. Five were serotyped, and the serovars Typhimurium and Anatum were associated with two cases each, while Muenster was associated with a case whose lesion pattern varied. Immunohistochemical staining was positive in all cases. All diagnoses were based on the clinical history, macroscopic and histological lesions, and the bacterial isolation and/or immunostaining associated with histological lesions.     

A study on some reproductive disorders in dromedary camel herds in Saudi Arabia with special references to uterine infections and abortion

Dromedary camels complaining from conception failure or abortion were investigated and their herders interviewed in Al Ahsa province, Kingdom of Saudi Arabia (KSA) during 2013 and 2015. The most important reproductive disorder according to the responders is uterine infection (60.2%) followed by obesity (22.3%) then physiological conditions (hormonal disturbances; 7.8%), adhesions (3.9%) and repeat breeders (2.9%). Of the camel herders, 78.6% reported previous occurrence of abortion in their herds and 46% reported abortion cases in the last season (2015/2016), while 21.4% reported no history of abortion. Most of the responders (97.1%) do not call a veterinarian for cases of abortion in their herds and 53.4% do not discard aborted materials. The majority of the herders (76.7%) deny that handling aborted materials or touching vaginal fluids can result in human infection, or replied they do not know. Uterine swab samples were collected and tested by PCR for seven potential pathogens and sera tested for antibodies against bovine viral diarrhea virus (BVDV) and Brucella. Five pathogens were identified in infected uterine samples, namely Coxiella burnetii (36%), Campylobacter spp. (27%), Brucella spp. (17%), Salmonella spp. (13%), and Chlamydia spp. (7%). Sero-prevalence of Brucella and BVDV was 8.2 and 29.1% in overall sera, respectively, and varies with regard to the region. The findings of the present study demonstrate that reproductive disorders dominated by uterine infections and abortions are widespread in dromedary camels in KSA.     

Protein requirements of collared peccary (<Emphasis Type="Italic">Pecari tajacu</Emphasis>)

A nitrogen (N) balance digestion trial was conducted to determine the protein requirement of collared peccaries (Pecari tajacu). In a 4 × 4 Latin square design, four captive adult male peccaries were fed four isoenergy diets containing four different levels of N (11.7, 16.3, 22.8, and 26.7 g N/kg of dry matter—DM). After 15 days of adaptation, a total collection of feces and urine was carried out for five consecutive days. Regression analyses between N intake and N in feces and urine allowed to calculate the metabolic fecal nitrogen (MFN = 2.3 g N/kg of dry matter intake—DMI) and daily endogenous urinary N (EUN = 185 mg N/kg^0.75). Likewise, by regression analyses between consumption of nitrogen and the nitrogen balance (NB = N ingested ? N excreted, mg N/kg^0.75), a daily requirement of 514 mg N/kg^0.75 was calculated. Therefore, if food intake is unrestricted, collared peccaries require a minimum in their diet of about 5.4% crude protein on DM basis. These values are almost as low as those found for browsing and frugivorous wild ruminants, which reinforce the proposition that peccaries’ digestive physiology is nearer to that of domestic and wild ruminants than domestic pigs. This relatively low protein requirement of collared peccary and its great ability to digest protein reveal the relevance of the forestomach for the species on nitrogen/protein metabolism and allow the use of diets with lower crude protein levels than the commercial ones used for the domestic pig, which reduces feed costs.     

Brucellosis in migratory sheep flock from Maharashtra,India

Brucellosis is a zoonotic disease worldwide distributed and having the economic as well as public health importance. The prevalence of brucellosis among sheep flock having history of abortions was studied. A total of 229 samples comprising of 157 blood and 72 clinical samples (vaginal swabs) were collected from 157 animals. Clinical samples were processed for the isolation of Brucella melitensis. Serum samples (n = 157) were tested by Rose Bengal plate test (RBPT) and i-ELISA. A total of 68 (43.31%) and 104 (66.24%) samples were positive by RBPT and ELISA, respectively. Brucella isolates (n = 2) were recovered from clinical samples. Both isolates demonstrated amplification for bcsp 31 and IS711 genes. On AMOS PCR, both the isolates amplified at 731 bp, i.e., belongs to B. melitensis species. The incidence of B. melitensis in a migratory flock warns the thorough testing and culling of Brucella-infected sheep from the flock on a continuous basis; otherwise, such incidence will be routine and poor farmers will be at a loss.     

The IMPDH inhibitors,ribavirin and mycophenolic acid,inhibit peste des petits ruminants virus infection

Peste des petits ruminants virus (PPRV) causes highly contagious diseases in domestic and particular wild small ruminants, leading to substantial economic loss. The development of effective and cheap antiviral medications shall help to circumvent this emerging burden. In this study, we found that ribavirin, a competitive inosine-5’-monophosphate dehydrogenase (IMPDH) inhibitor, significantly inhibits the replication of PPRV. As IMPDH is a key enzyme in purine nucleotide synthesis, supplementation of exogenous guanosine attenuate the anti-PPRV effect of ribavirin. Interestingly, an uncompetitive IMPDH inhibitor, mycophenolic acid (MPA), exerted more potent antiviral effect again PPRV. Similarly, this effect was largely restored upon supplementation of guanosine. Thus, we have demonstrated that the IMPDH inhibitors ribavirin and MPA combat PPRV infection through purine nucleotide depletion. Because both regimens have been widely used in the clinic for treating viral infection or organ rejection in transplantation patients for decades, respectively, repurposing these existing safe and cheap medications may provide a new avenue for combating PPRV infection.