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.     

Estimates of direct and maternal (co)variance components as well as genetic parameters of growth traits in Nellore sheep

In the present study, (co)variance components and genetic parameters in Nellore sheep were obtained by restricted maximum likelihood (REML) method using six different animal models with various combinations of direct and maternal genetic effects for birth weight (BW), weaning weight (WW), 6-month weight (6MW), 9-month weight (9MW) and 12-month weight (YW). Evaluated records of 2075 lambs descended from 69 sires and 478 dams over a period of 8 years (2007–2014) were collected from the Livestock Research Station, Palamaner, India. Lambing year, sex of lamb, season of lambing and parity of dam were the fixed effects in the model, and ewe weight was used as a covariate. Best model for each trait was determined by log-likelihood ratio test. Direct heritability for BW, WW, 6MW, 9MW and YW were 0.08, 0.03, 0.12, 0.16 and 0.10, respectively, and their corresponding maternal heritabilities were 0.07, 0.10, 0.09, 0.08 and 0.11. The proportions of maternal permanent environment variance to phenotypic variance (Pe²) were 0.07, 0.10, 0.07, 0.06 and 0.10 for BW, WW, 6MW, 9MW and YW, respectively. The estimates of direct genetic correlations among the growth traits were positive and ranged from 0.44(BW-WW) to 0.96(YW-9MW), and the estimates of phenotypic and environmental correlations were found to be lower than those of genetic correlations. Exclusion of maternal effects in the model resulted in biased estimates of genetic parameters in Nellore sheep. Hence, to implement optimum breeding strategies for improvement of traits in Nellore sheep, maternal effects should be considered.     

Relationship between teat morphological traits and subclinical mastitis in Frieswal dairy cows

The present study was aimed to investigate the relationship between selected morphological traits of teat and subclinical mastitis (SCM) in Frieswal crossbred dairy cows. A total of 1040 quarters from 261 lactating cows were evaluated for teat shape (bottle/fleshy/collapsed/conical/normal/pencil and short), teat-end shape (dished/flat/funnel/pocketed/pointed and rounded), teat orientation (aligned/misaligned) and teat position (front and rear; left-sided and right-sided). Each udder quarter was screened with California Mastitis Test (CMT) for the purpose of defining quarter health status. Data were analysed using Chi-square test and multivariable logistic regression procedure. An overall prevalence of SCM (CMT positive) at quarter level was 30.6%. Most of the teats had normal or cylindrical shapes (48%), dished teat-ends (40.7%), and aligned (central or squared) in orientation (65%). At bivariable level, significant association of SCM with teat shape, teat position, teat orientation, parity, and stage of lactation was observed (P < 0.05 to P < 0.001). Teat-end shapes showed some association with SCM (P = 0.07). Results of multivariable analysis showed that pencil-shaped teats were least associated with SCM (P < 0.05) as compared to other teat shapes. Prevalence of SCM was also higher in rear teats (P = 0.015), misaligned teats (P = 0.01), and cows in second or higher parities (P < 0.01) and late stage of lactation (P < 0.001). The results of the present study indicate that selected morphological traits of teat are associated with SCM in Frieswal crossbred cows; therefore, selection towards desirable morphological traits could help reduce mastitis in this breed.     

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.     

Use of serology and real time PCR to control an outbreak of bovine brucellosis at a dairy cattle farm in the Nile Delta region,Egypt

Background

Bovine brucellosis remains one of the most prevalent zoonotic infections affecting dairy cattle in developing countries where the applied control programs often fail. We analyzed the epidemiologic pattern of bovine brucellosis in a dairy cattle herd that showed several cases of abortions after regular vaccination with RB51 (B. abortus vaccine). In 2013 thirty dairy cows, from a Holstein-Friesian dairy herd with a population of 600 cattle, aborted five months post vaccination by a regular RB51 vaccine. Blood samples were drawn from milking cows and growing heifers, as well as heifers and cows pregnant up to 6 months. These samples were collected in June 2013 (n?=?257) and May 2014 (n?=?263) and were tested by real time (rt)-PCR as well as serological tests, in particular Rose Bengal Test (RBT), Enzyme-Linked Immunosorbent Assays (ELISA) and Fluorescence Polarization Assay. Tissue specimens were also collected from an aborted fetus and cultured. Isolates were subjected to bacteriological typing tests at the genus and species levels.

Results

Five months post vaccination with RB51 vaccine, Brucella (B.) DNA was detected in blood samples of cows by rt-PCR. The serological tests also revealed the spread of Brucella field strains within the herd in 2013. Four Brucella isolates were recovered from specimens collected from the aborted fetus. These isolates were typed as follows: one B. abortus RB51 vaccine strain and three isolates of B. abortus field strain. The seropositive cows with positive rt-PCR might indicate an infection by the Brucella field strain; while the positive rt-PCR results from seronegative animals may either be due to circulating RB51 vaccine DNA in vaccinated animals or to circulating field strain in infected animals before seroconversion.

Conclusion

The results herein suggest that PCR can be a good supplementary tool in an outbreak situation, if an assay is available that can differentiate vaccine and field strains with a high analytical sensitivity. We recommend using RBT and ELISA in parallel in outbreak situations, to identify as many infected animals as possible during the initial screenings. This test procedure should be repeated for at least three successive negative tests, with one month interval.

     

Canine fetal echocardiography: correlations for the analysis of cardiac dimensions

The aim of this study was to develop regression models for correlation of canine fetal heart development with body size to characterize normal development or suggest cardiac anomalies. Twenty clinically healthy pregnant bitches, either brachycephalic and non-brachycephalic, were examined ultrasonographically. Transabdominal fetal echocardiography was conducted every 4 days from the beginning of cardiac chambers differentiation until parturition. Ten cardiac parameters were measured: length, width and diameter of the heart; heart area; left and right ventricular dimensions; left and right atrial dimensions; and aortic and pulmonary artery diameter. Femoral length, biparietal diameter and abdominal cross-sectional area were also recorded. Regression equations were developed for each parameter of fetal body size, and linear and logarithmic models were compared. The model with the highest correlation coefficient was chosen to produce equations to calculate relative dimensions based on the correlations. Only the left-ventricular chamber differed between the two racial groups. Biparietal diameter was the independent parameter that produced the highest correlation coefficient for the most fetal cardiac dimensions, although good correlations were also observed using femoral length and abdominal cross-sectional area. Heart width and heart diameter were used as surrogates of cardiac development, as these measurements showed the best statistical correlation. Quantitative evaluation of fetal cardiac structures can be used to monitor normal and abnormal cardiac development.     

Long-term effects of good handling practices during the pre-weaning period of crossbred dairy heifer calves

The aim of this study was to determine whether applying good practices of handling during the pre-weaning period have long-term effects on behavioral and physiological indicators, health status, and average daily gain (ADG) of crossbred Bos taurus × Bos indicus heifer calves. During the pre-weaning period, 98 crossbred of Holstein × Gir heifer calves were allotted into three treatments: (1) good practices of handling + brushing (GPB; n?=?25), (2) good practices of handling (GP; n?=?25), and (3) control (n?=?48). Every 2 months, four evaluation periods (EV₁ to EV₄) were conducted to record data. Behavioral indicators comprised time to drive (TD), flight speed (FS), flight distance (FD), and composite reactivity score (CRS). Physiological indicators of acute stress during handling comprised respiratory and heart rates. Health status comprised data regarding occurrence of most common diseases (i.e., pneumonia and anaplasmosis). Collected data were analyzed by using a linear mixed model for repeated measures, Tukey’s test, and chi-squared procedures. Treatment influenced (P?<?0.05) TD, FS, and FD but not CRS (P?=?0.78). From EV₁ to EV₃, the control calves had the lowest TD. The GPB group had lower FS than the control but did not differ from GP. The GPB group had lower FD means than the other two groups in EV₂, EV₃, and EV₄. No differences (P?>?0.05) due to treatment were observed on heart and respiratory rates, ADG, or occurrence of pneumonia and anaplasmosis. It was concluded that adoption of good practices of handling during pre-weaning period may lead to long-term positive effects.     

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).