An assessment of benzimidazole resistance against caprine nematodes in Central India

Current status of resistance to benzimidazole (BZ) group of anthelmintic drugs against caprine nematodes in Central India at Amanala goat farm, Jabalpur, Madhya Pradesh (M. P.), was systematically investigated using faecal egg count reduction (FECR) test and egg hatch test (EHT). Besides, allele-specific PCR (AS-PCR) was deployed to ascertain the susceptible genotype (alleles) especially of the Haemonchus contortus. Randomly selected 30 goats, irrespective of age and sex, were divided into three groups of 10 each, to serve as treated and untreated controls. It was ensured that the animals were not administered with an anthelmintic drug for the past 3 months prior to undertaking the study, and faecal egg counts were estimated. FECR test evidenced fenbendazole resistance by partial elimination (24.90%) copro-egg counts in the treated group of animals vis-à-vis controls with a lower confidence interval of ?26%. Further, EHT revealed ED-50 value of 0.335 μg of thiabendazole/ml, confirming benzimidazole resistance in the animals of that farm. AS-PCR showed that 62% of H. contortus larvae were homozygous resistant (rr), 24% heterozygous (rS) and 14% homozygous susceptible (SS). The genotypic frequencies of three genotypes (rr, rS and SS) were significantly (P < 0.01) different. The prevalence of benzimidazole resistance allele (r) was also significantly (P < 0.01) higher (74%) as compared to susceptible allele (S) (26%). The resistance to benzimidazole has been discussed while emphasizing improved managemental practices designed to reduce exposure of the goat population to parasites, minimize frequency of anthelmintic use at optimum dose and rotational use of different chemical groups of medicines with different mode of action, so as to overcome and combat the upcoming problem in the field.     

Growth,feed efficiency and blood profile of buffalo calves consuming high levels of fluoride

Twelve male buffalo calves of 10 to 12 months of age were divided into 3 groups of four each. They were fed wheat straw+concentrate mixture +3 Kg greens. The chemical composition of the diet was same in all the three groups except fluoride which was added (as NaF) in concentrate mixture of group B and C to make the final fluoride concentration 30 ppm and 60 ppm respectively. The animals were kept on scheduled diet for a period of 90 days. Body weights were recorded at the start of the experiment and at fortnightly interval thereafter. Analysis of data revealed that the dry matter intake decreased non significantly in group B and C as compared to control group. A significant decrease in serum calcium and a significant increase in phosphorus concentration were observed in group C animals. A significant increase was observed in alkaline phosphatase activity in group C animals. A non significant decrease was observed in T₄ values in group C animals. On the basis of these results it could be concluded that fluoride in the diet of buffalo calves @ 30 ppm is a safe level whereas 60 ppm has affected the blood metabolites.     

Development of a murine ocular posterior segment explant culture for the study of intravitreous vector delivery

The objective of this study was to develop a murine retinal/choroidal/scleral explant culture system to facilitate the intravitreous delivery of vectors. Posterior segment explants from adult mice of 2 different age groups (4 wk and 15 wk) were cultured in serum-free medium for variable time periods. Tissue viability was assessed by gross morphology, cell survival quantification, activated caspase-3 expression, and immunohistochemistry. To model ocular gene therapy, explants were exposed to varying transducing units of a lentiviral vector expressing the gene for green fluorescent protein for 48 h. Explant retinal cells remained viable for approximately 1 wk, although the ganglion cell layer developed apoptosis between 4 and 7 d. Following vector infusion into the posterior segment cups, viral transduction was noted in multiple retinal layers in both age groups. An age of donor mouse influence was noted and older mice did not transduce as well as younger mice. This explant offers an easily managed posterior segment ocular culture with minimum disturbance of the tissue, and may be useful for investigating methods of enhancing retinal gene therapy under controlled conditions.     

Validating a Time Series of Annual Grass Percent Cover in the Sagebrush Ecosystem

We mapped yearly (2000–2016) estimates of annual grass percent cover for much of the sagebrush ecosystem of the western United States using remotely sensed, climate, and geophysical data in regression-tree models. Annual grasses senesce and cure by early summer and then become beds of fine fuel that easily ignite and spread fire through rangeland systems. Our annual maps estimate the extent of these fuels and can serve as a tool to assist land managers and scientists in understanding the ecosystem’s response to weather variations, disturbances, and management. Validating the time series of annual maps is important for determining the usefulness of the data. To validate these maps, we compare Bureau of Land Management Assessment Inventory and Monitoring (AIM) data to mapped estimates and use a leave-one-out spatial assessment technique that is effective for validating maps that cover broad geographical extents. We hypothesize that the time series of annual maps exhibits high spatiotemporal variability because precipitation is highly variable in arid and semiarid environments where sagebrush is native, and invasive annual grasses respond to precipitation. The remotely sensed data that help drive our regression-tree model effectively measures annual grasses’ response to precipitation. The mean absolute error (MAE) rate varied depending on the validation data and technique used for comparison. The AIM plot data and our maps had substantial spatial incongruence, but despite this, the MAE rate for the assessment equaled 12.62%. The leave-one-out accuracy assessment had an MAE of 8.43%. We quantified bias, and bias was more substantial at higher percent cover. These annual maps can help management identify actions that may alleviate the current cycle of invasive grasses because it enables the assessment of the variability of annual grass ? percent cover distribution through space and time, as part of dynamic systems rather than static systems.     

Level and period of realimentation to assess improvement in body condition and carcass quality in cull ewes

Improvement in body condition was assessed in 40 cull ewes (>6 years), equally distributed in two groups and realimented with ad libitum roughage (gram straw) and two levels of concentrate feeding, i.e., 2.5 % (T₁) of live weight (LW) and ad libitum (T₂). Five representative animals from an initial 45 were slaughtered at the initiation of the study (0 day) and five animals from each treatment at 44, 67, and 90 days of experiment for carcass attributes. Improvement in body condition score (BCS), nutrient utilization, feed efficiency, and carcass traits were assessed at 44, 67, and 90 days. Metabolism trial of 6-day collection of feed, feces, and urine samples was conducted on five representative ewes from each group after 60 days of feeding. The level of concentrate feeding on LW gain and BCS was significant, and the duration of realimentation showed a linear improvement (P?<?0.001). The digestibility and intake of dry matter, organic matter, and crude protein was higher (P?<?0.05) in T₂. The N intake, absorption, and balance showed a similar trend. Increase (P?<?0.05) in total N, trichloroacetic acid precipitable N, and ammonia N was observed with extension of realimentation period. Blood metabolic profile also showed improvement (P?<?0.05) from an undernourished state to normal after alimentation. Animals in T₂ accumulated higher LW with minimal expenditure of metabolizable energy (73.4 vs 79.1 MJ) and higher efficiency of feed conversion during 68 to 90 days of realimentation. Linear improvement (P?<?0.01) in carcass traits (preslaughter weight, empty live weight, hot carcass weight, dressing percentage, and amount of subcutaneous and intramuscular fat) and composition of longissimus dorsi muscle was observed. Ad lib concentrate supplementation for a period of 90 days may thus be considered appropriate for achieving desired efficiency of gain and improvement in body condition of cull ewes for quality mutton production.     

Exogenous DNA internalisation by sperm cells is improved by combining lipofection and restriction enzyme mediated integration

1. Three types of exogenous DNA inserts, i.e. complete linearised pVIVO2-GFP/LacZ vector (9620 bp), the LacZ gene (5317 bp) and the GFP gene (2152 bp) were used to transfect chicken spermatozoa through simple incubation of sperm cells with insert. 2. PCR assay, Dot Blot hybridisation and Southern hybridisation showed the successful internalisation of exogenous DNA by chicken sperm cells. 3. Lipofection and Restriction Enzyme Mediated Integration (REMI) were used to improve the rate of internalisation of exogenous DNA by sperm cells. 4. Results from dot blot as well as Southern hybridisation were semi-quantified and improved exogenous DNA uptake by sperm cells through lipofection and REMI. Stronger signals were observed from hybridisation of LacZ as well as GFP specific probe with the DNA from lipofected exogenous DNA transfected sperm DNA in comparison with those transfected with nude exogenous DNA.     

Fertility following CIDR based synchronization regimens in anoestrous Nili-Ravi buffaloes

The objective of this study was to compare oestrus expression and fertility rate in used and new controlled internal drug releasing (CIDR) device treated anoestrous buffaloes. Furthermore, to determine the timing of ovulation, and fertility rate in estradiol benzoate (EB) and GnRH-administered CIDR-treated anoestrous Nili-Ravi buffaloes. In experiment 1, buffaloes received either a used CIDR (UCIDR, n = 35) or a new CIDR (NCIDR, n = 36) for 7 day and PGF2α on day 6. Oestrous expression was similar (p > 0.05) between UCIDR (88.5%) and NCIDR (96.6%) buffaloes. The pregnancy rate did not differ (p > 0.05) because of treatment (37.1% in UCIDR vs 36.6% in NCIDR). In experiment 2, buffaloes (n = 55) received CIDR device for 7 days and PGF2α, on day 6 and randomly assigned into three treatment groups: (i) CIDR-EB (n = 17) received EB on day 8, (ii) CIDR-GnRH (n = 18) received GnRH on day 9 and (iii) control (n = 20) received no further treatment. Mean interval from CIDR removal to ovulation in CIDR-EB, CIDR-GnRH and CIDR group were 61.3 ± 0.8, 64.9 ± 1.8 and 65.1 ± 16.7 h, respectively. However, the buffaloes in the CIDR-EB and CIDR-GnRH group had lesser variability in the timing of ovulation compared to control. The pregnancy rate of both CIDR-EB group (58%) and CIDR-GnRH group (61%) were tended to be higher (p < 0.1) than control (30%). In conclusion, compared to NCIDR devices, previously UCIDR devices are equally effective to induce oestrus in anoestrous buffaloes resulting optimal pregnancy rate. Administration of EB and GnRH after CIDR removal results in tighter synchrony (less variability) and improved fertility in anoestrous buffaloes. CIDR based synchronization regimens have great potential in fertility improvement in anoestrous buffaloes.     

Toxicological effects and recovery of the corneal epithelium in Cyprinus carpio communis Linn. exposed to monocrotophos: an scanning electron microscope study

This study was conducted based on the evidence of fish habitats in North India being affected by organophosphate pesticides draining from agricultural fields into bodies of water, especially during the rainy season. Various tissues of fish such as scales, gills ovaries, kidney, and liver have been studied from the toxicological point of view, but the toxicological effects of aquatic pollutants on fish cornea have not been investigated to date. We conducted comparative toxicological studies on the cornea of Cyprinus carpio communis using two sublethal (0.038 and 0.126 ppm) concentrations of monocrotophos pesticide for 30 days. Corneas from all the groups were evaluated by a scanning electron microscope. The fish exposed to the monocrotophos pesticide developed corneal necrosis due to the formation of crystalloid‐like structures, thinning and shrinkage of microridges on the corneal epithelium. After 30 days, fish from the monocrotophos‐treated tank were transferred to normal environmental conditions. After 60 days under natural condition, epithelial cells did not fully recover. In conclusion, exposure to monocrotophos induces irreversible changes in the cornea of C. carpio communis. As fish and mammalian visual systems share many similarities, the reported finding may offer useful insights for further toxicological and ophthalmological studies in humans.