Physio-biochemical parameters: a potential tool for target-selective treatment of haemonchosis in the small ruminants

This study aims to evaluate the conjunctiva colour-based FAMACHA score (FS) coupled with a body condition score (BCS), haemogram and stressor hormone level estimation, in identifying post-mortem (PM)/coproscopically proven individuals wanting therapy for economically important gastrointestinal (GI) helminths, Haemonchus contortus, in the small ruminants. The incidence of haemonchosis was significantly (p < 0.05) higher (60.81%) in the ruminants with FS = 3. The H. contortus count in the animals with FS 2, 3 and 4 was 23.2 ± 0.37, 62 ± 2.5 and 74 ± 3.2 (p < 0.05) [positive correlation (r = 0.841 in goats; r = 0.828 in sheep, p < 0.05)], respectively, with corresponding 2.8 ± 0.15, 2 ± 0.3 and 2 ± 0.16 BCS (negative correlation, p > 0.05). The infected animals of FS 2, 3 and 4 measured 8.2 ± 0.0, 7.5 ± 0.23 and 6.7 ± 0.34 g/dl Hb (r = ?0.452, p = 0.01) in goats/9.3 ± 0.8, 8.6 ± 0.5 and 7.6 ± 0.3 g/dl Hb (r = ?0.511, p = 0.05) in sheep with 21.2, 19.8 ± 1.8 and 17.8 ± 0.2% PCV (r = ?0.369, p = 0.05) in goats/26.7 ± 1.2, 22.2 ± 0.2 and 20.9 ± 0.6% PCV (r = ?0.251, p = 0.03) in sheep, respectively. The FS 2, 3 and 4 infected goats/sheep measured 6.1 ± 0, 7.9 ± 1.0 and 9.5 ± 0.9 (p < 0.05)/5.8 ± 2.3, 6.9 ± 1.2 and 7.8 ± 0.2% (p < 0.05) mid-granulocyte [(r = 0.928 (goats)/0.834 (sheep), p < 0.05], while the cortisol level was 15.6, 23 ± 4.5 and 42 ± 2.3 (p = 0.23)/12.1 ± 0, 15.9 ± 1.2 and 24 ± 3.4 (p = 0.29) μg/dl, respectively. The infected ruminants recorded low (p < 0.05) level of Hb/PCV while high level of mid-granulocytes/cortisol. Specificity of FAMACHA test was maximized (100%) when FS = 4 was considered anaemic, but sensitivity was low (35.29% in goats; 25% in sheep). The false negatives was 5.9 (goat)/12.5 (sheep)% when FS ≥ 3 was considered anaemic. The small ruminants with FS ≥ 3, BCS ≤ 2.5, Hb ≤ 7.5 g/dl (goats)/8.6 g/dl (sheep), PCV ≤ 19.8% (goats)/22.2% (sheep) and mid-granulocyte ≥7.9% (goats)/6.9 ± 1.2% (sheep) can be subjected to target-selective treatment for haemonchosis in the field simultaneously maximizing the economic benefit to the farmers.     

Molecular characterization of Ascaridia galli from Bangladesh and development of a PCR method for distinguishing A. galli from Heterakis spp.

We analyzed the nuclear ribosomal internal transcribed spacer (ITS) 1 and ITS2 sequences for Bangladesh isolates of Ascaridia galli, and we determined that the sequences were unreliable as molecular markers for distinguishing A. galli from other Ascaridia species, because the sequences showed high identity with that of A. columbae. However, the ITS1 sequences were available for designing PCR primers distinguishable between Ascaridia galli and Heterakis spp. Bangladesh isolates of A. galli constituted a monophyletic clade along with other geographical isolates in the cytochrome c oxidase subunit I (COI) phylogenetic tree, however, we could not clarify the phylogenetic relationships between A. galli and other Ascaridia spp., because their available sequences in GenBank were very few. The developed PCR method using DNA from A. galli and Heterakis spp. eggs would enable differential diagnosis of the individual infections in the future.     

Comparative morphometrical studies on the skull bones of barking deer (Muntiacus muntjak) and sambar deer (Rusa unicolor)

The present study reports data on the skull bone morphometry of barking and sambar deer. The skulls of adult barking deer (n = 6) and sambar deer (n = 6) of either sex (n = 3 males and n = 3 females) were collected from the Aizawl Zoological Park, Aizawl, Mizoram, India, with official permission from the Government of Mizoram. Anatomically, barking and sambar deer's skulls were elongated, pyramid-like, dolichocephalic and consisted of thirty-two cranial and facial bones. The cranial bones were eleven (three single and four paired), comprising of occipital, sphenoid, ethmoid, frontal, interparietal, parietal and temporal. The facial bones were twenty-one (one single and ten were paired), consisting of the maxilla, premaxilla (incisive), palatine, pterygoid, nasal, lacrimal, zygomatic (malar), vomer, turbinates, mandible and hyoid. In the present study, altogether 41 different measurements were taken morphologically and 6 different indices were applied. The obtained morphometrical parameters were significantly (p < .01, p < .05) higher in males than females of both species. Species wise, all obtained parameters were higher in sambar deer than barking deer. The obtained 41 different skull parameters and 6 indices showed statistically significant differences (p < .01 and p < .05) between both sexes of barking and sambar deer; however, practically these differences were meagre. The present morphometrical study on the skull of both species can help the wildlife professionals and zoo veterinarians determine the sex of these animals and differentiate it from other domestic and wild small ruminants for solving veterolegal cases. This study's findings will also motivate and assist other comparative studies with various domestic and wild small ruminants.     

Detection and molecular characterization of naturally transmitted sheep associated malignant catarrhal fever in cattle in India

Malignant catarrhal fever (MCF) is a fatal herpesvirus infection of domestic and wild ruminants, with a short and dramatic clinical course characterized primarily by high fever, severe depression, swollen lymph nodes, salivation, diarrhea, dermatitis, neurological disorders, and ocular lesions often leading to blindness. In the present study, fatal clinical cases of sheep associated malignant catarrhal fever (SA-MCF) were identified in cattle in the state of Karnataka. These cases were initially presented with symptoms of diarrhea, respiratory distress, conjunctivitis, and nasal discharges. Laboratory diagnosis confirmed the detection of ovine herpesvirus-2 (OvHV-2) genome in the peripheral blood samples of two ailing animals. The blood samples collected subsequently from sheep of the neighboring areas also showed presence of OvHV-2 genome indicating a nidus of infection in the region. The positive test results were further confirmed by nucleotide sequencing of the OIE approved portion of tegument gene as well as complete ORF8 region of the OvHV-2 genome. Phylogenetic analysis based on the sequence of the latter region indicated close genetic relationship with other OvHV-2 reported elsewhere in the world.     

Effect of Incorporation of Additives in Tris‐Based Egg Yolk Extender on Buffalo (Bubalus bubalis) Sperm Tyrosine Phosphorylation During Cryopreservation

Phosphorylation of tyrosine residues on sperm protein is a known indicator of capacitation and a major intracellular signalling event. There is evidence that sperm cryopreservation promotes tyrosine phosphorylation and is associated with reduced fertility of spermatozoa. Under this study, cryoprotective role of different additives namely taurine, trehalose, catalase and 4‐bromophenacyl bromide on buffalo sperm quality was evaluated. Buffalo semen was cryopreserved in tris‐based egg yolk extender supplemented with additives like taurine (50 mm ) or trehalose (100 mm ) or 4‐bromophenacyl bromide (200 μm ) or catalase (100 U/ml) and used for assessment of levels of tyrosine phosphorylation in frozen‐thawed spermatozoa. The results obtained were compared with the level of protein tyrosine phosphorylation of semen cryopreserved in tris‐based egg yolk extender without additives. Proteins were extracted from a total number of nine ejaculates from three individual buffalo bulls chosen at random and analysed for tyrosine phospho‐proteins using SDS–PAGE followed by immunoblotting. Monoclonal anti‐phosphotyrosine antibody (Clone pT‐154) was used as primary antibody followed by treatment with HRP‐conjugated secondary antibody. Signals were detected on X‐ray film using chemiluminescence. Nine proteins (p20, p30, p32, p38, p49, p56, p59, p72 and p86) were found to be tyrosine phosphorylated in cryopreserved spermatozoa. Supplementation of additives significantly (p<0.05) reduced the level of protein tyrosine phosphorylation in spermatozoa. Moreover, this study showed improved (p<0.05) post‐thaw motility, viability and membrane integrity of spermatozoa on addition of these additives. The results obtained clearly indicate reduced level of capacitation like changes on supplementation of additives in terms of protein tyrosine phosphorylation.     

Larval and pupal toxicity effects of Plectranthus amboinicus, Bacillus sphaericus and predatory copepods for the control of the dengue vector, Aedes aegypti

The efficacy of copepod Mesocyclops aspericornis (Daday) combined with the larvicide Bacillus sphaericus (Bs) and a plant extract of Plectranthus amboinicus leaf abstract (PALE), used jointly and singly, was studied against Aedes aegypti in the laboratory. P. amboinicus leaf extract of 20, 40, 60, 80 and 100 ppm caused significant mortality against Ae. aegypti larvae. The LC₅₀ and LC₉₀ values for I to IV instars larvae and pupae were 26.12, 35.36, 45.76, 52.32 and 63.82 ppm, respectively. The LC₉₀ values for I to IV instars larvae and pupae were 82.53, 92.65, 108.06, 119.47 and 131.71 ppm, respectively. Under laboratory conditions, copepods treatment produced 7.9% predatory efficiency against 1st instar larvae of Ae. aegypti, at a copepod:larvae ratio of 1:10. When copepod treatment was combined with PALE this was increased to 8.7. The treatment of copepods combined with Bs and PALE yielded a better and more sustainable result (9.6%) than the agents used individually. This predation efficiency may be caused by detrimental effects of the P. amboinicus active principle compound (carvacrol) on the mosquito larvae. Our results suggest that the combined application of microbial insecticide (Bs), copepods and P. amboinicus leaf extract may be used to control Aedes populations.