In vitro maturation and fertilization in the Nilgai (Boselaphus tragocamelus) using oocytes and spermatozoa recovered post-mortem from animals that had died because of foot and mouth disease outbreak

The ability to rescue gametes from endangered or wildlife species and to subsequently produce viable embryos holds tremendous potential as a means to increase the population size of endangered or wildlife species. The objective of this study was to assess the developmental competence of gametes recovered from nilgai that had died because of foot and mouth disease outbreak. Oocytes collected from the ovaries of seven dead nilgais were allowed to mature in vitro and were tested for developmental potential by in vitro fertilization (IVF) with epididymal spermatozoa collected also post-mortem. The average number of oocytes (n = 517) recovered per ovary was 36.9, and the side (right or left), size and weight of the ovaries had no significant effect on the number and quality of oocytes recovered. In vitro maturation studies indicated that the proportion of matured oocytes (MII stage) at 18, 24 and 30 h was 55.6%, 63.4% and 63.6%, respectively. Furthermore, 43% of the matured oocytes cleaved following in vitro fertilization and 12% of the cleaved oocytes (6/49) developed to the 4-8 cell stage. These findings suggest that the gametes recovered from nilgai post-mortem could be utilized for in vitro production of embryos.     

Increased androgen receptor expression in muscle tissue contributing to growth increase in androgen-treated Nile tilapia

The aim of this study was to investigate the molecular mechanism behind androgen-induced sex reversal and growth increase in Nile tilapia. Three-day-old mixed-sex tilapia juveniles were treated with differential dose (0, 5, 10, 20, 30 mg/kg diet) and duration (0, 15, 30, 45, 60 days) regimes of 17α-methyltestosterone. Treatment regimes 10, 20, 30 mg/kg-30, 45, 60 days showed ~95% males, while 0 mg/kg-0 days (control) showed ~50% males (P < 0.01). Next, tilapia was cultured for 6 months in cistern and pond culture systems with 17α-methyltestosterone-treated diet (10 mg/kg) for three different durations: 0 days (control), 30 days (30-day-treated), and 180 days (throughout-treated). Fish in pond culture showed significantly higher (P < 0.05) final weight compared with tilapia in cistern culture (235.76, 156.82 g, respectively). Both 30-day-treated and throughout-treated fish showed significantly higher (P < 0.05) final weight, length, daily weight gain, specific growth rate, and muscle protein content compared with control fish. The androgen receptor β mRNA and protein expression levels in both 30-day-treated and throughout-treated fish were significantly higher (~twofolds, P < 0.05) compared with hormone-untreated control males. However, there was not much difference for all these parameters among the two hormone-treated groups. This higher growth probably results from an androgen-induced increased ‘androgen-androgen receptor’ mediated anabolic pathway. Serum testosterone concentration of 30-day-treated fish after 6 months was similar to that of control males (0.9 ng/ml). Moreover, culture of 17α-methyltestosterone-treated monosex tilapia was found beneficial than hormone-untreated mixed-sex tilapia culture for maintenance of natural fish diversity in indigenous water bodies.     

Evaluation of safety and efficacy of acaricides for control of the African tortoise tick (Amblyomma marmoreum) on leopard tortoises (Geochelone pardalis).

Nine acaricides (amitraz, carbaryl, chlorpyrifos, cyfluthrin, fipronil, lindane, permethrin, phenothrin, and pyrethrins) were studied for their efficacy in killing the African tortoise tick (Amblyomma marmoreum). Only four of the acaricides (chlorpyrifos, cyfluthrin, lindane, and permethrin) produced 100% mortality within 24 hr of application, and only two (cyfluthrin and permethrin) continued to cause 100% mortality when diluted to as low as 1:10,000. Five of the acaricides (amitraz, carbaryl, chlorpyrifos, cyfluthrin, and permethrin) were studied for toxicity to the leopard tortoise (Geochelone pardalis), the most common host of A. marmoreum. The results indicate that cyfluthrin and permethrin, which were herein found to be the two most effective acaricides for control of A. marmoreum, are also the safest acaricides for use on leopard tortoises.     

Evaluation and impact assessment of culture‐based fisheries to enhance fish yield in small reservoirs in Odisha State,India

The present study investigates the success of stocking fingerlings of Indian major carps ‐Gibelion catla (Hamilton), Labeo rohita Hamilton and Cirrhinus mrigala Hamilton – to enhance fish yield in 58 small reservoirs in Odisha State, India. Fish stocking and yield data were collected from State Fisheries Department, Odisha. The relationships between area, stocking density and fish yield of different size groups of reservoirs were assessed using the Pearson correlation coefficient (r). Fish yield increased significantly (p < 0.05, t‐test) from 204 kg/ha/year in 2012–13 to 323 kg/ha/year in 2013–14 due to adoption of CBF. The corresponding per capita fish production increased from 398 kg fisher^?1 year^?1 to 702 kg fisher^?1 year^?1 benefitting more than 8,000 fisher households. Reservoirs in the size group 100–500 ha showed the highest response of fish yield with respect to stocking of fingerlings. The size of fingerlings at stocking and area of the reservoir had greater impact on enhancing fish yield. Stocking had a profound positive impact on fish yield. This study forms a baseline evaluation of the impact of stocking on small tropical reservoirs in India. It also recommends management measures and discusses issues, and the way forward for sustainable fisheries enhancement in small tropical reservoirs in eastern India.     

Compatibility of silver barb Puntius gonionotus (Bleeker) with Indian major carps in a grow-out polyculture

The compatibility and growth performance of silver barb Puntius gonionotus (Barbonymous gonionotus) with the three Indian major carps, i.e., Catla catla, Labeo rohita and Cirrhinus mrigala, were assessed in a 10‐month carp polyculture trial. Treatments T‐1, T‐2, T‐3 and T‐4 were stocked with three of the above four carp species, with an absence of silver barb, mrigal, rohu and catla, respectively, while all four species were stocked in treatment T‐5. The treatments were stocked at 6000 fingerlings ha⁻¹, with an equal species ratio maintained in each treatment. Incorporation of silver barb into the polyculture system neither affected the survival of any carp irrespective of species combination nor yielded significant changes in biomass production among treatments, except for the one without catla, where it was significantly low. The study revealed a higher extent of competition between silver barb and rohu, perceptible from the lower growth of one in the presence of the other. Although a certain level of competition of silver barb with mrigal was evident, competition with catla was not perceptible. Irrespective of species combination with silver barb as a component species, similar total biomass production in treatments revealed the feasibility of its incorporation into the Indian major carp‐based polyculture practice without affecting the total yield.     

Detection and characterization of a phytoplasma associated with witches’-broom disease of Salvadora persica in India

Salvadora persica trees with symptoms of witches’-broom, little leaf and severe leaf curling were observed in the Biodiversity Park, North Delhi (India) during February 2010. Using a nested-PCR assay targeting two phytoplasma specific loci (16S ribosomal RNA and the ribosomal protein gene) and sequence analysis of the amplified products, we identified the associated phytopathogen as a phytoplasma of group 16SrI. RFLP-mediated characterization revealed that the phytoplasma belongs to subgroup 16SrI-B. Phylogenetic reconstruction also grouped the phytoplasma within group 16SrI. The study reports a new host of phytoplasma in India and worldwide.     

Increasing prevalence of Coxiella burnetii seropositive Danish dairy cattle herds

A study based on bulk tank milk samples from 120 randomly selected dairy cattle herds was conducted to estimate the prevalence of Coxiella burnetii seropositive dairy herds, to describe the geographical distribution, and to identify risk factors. Using the CHEKIT Q-fever Antibody ELISA Test Kit (IDEXX), the study revealed a prevalence of 79.2% seropositive herds, 18.3% seronegative herds, and 2.5% serointermediate herds based on the instructions provided by the manufacturer. Multifactorial logistic regression showed statistically significant associations (P < 0.01) between C. burnetii seropositivity and increasing herd size (OR = 1.02 per cow increment) and increasing regional average number of cattle per dairy herd (OR = 1.02 per animal increment). Herds >150 cows had 17.9 times higher odds of testing positive compared to herds <80 cows. The regional average number of cattle herds per square kilometer was borderline significantly related to the occurrence of seropositive dairy herds (P = 0.06). The results indicate an increased prevalence of seropositive dairy herds since the previous survey in 2008 and an adverse impact of increasing herd size and cattle density on the risk of seropositivity.     

Evaluation of promoters and visual markers for transformation of eastern white pine

This report serves to evaluate possible promoters for use in theproduction of transgenic eastern white pine (Pinus strobus L.).Embryogenic cultures of eastern white pine were bombarded with goldparticles coated separately with a variety of gene constructs containingthe UidA -glucuronidase (GUS) or green fluorescentprotein (GFP) reporter gene. Transient expression of the UidAgene, driven by a novel algal virus adenine methyl transferase genepromoter, as well as five other promoters used in angiospermtransformation, were evaluated. The maize alcohol dehydrogenase promoterwas not effective in eastern white pine cultures. The construct with thedoubled Cauliflower Mosaic Virus 35S promoter plus Alfalfa Mosaic Virusenhancer showed the highest levels of expression. GUS expression wasdetected within 24 hours, but decreased after 5 days and was notdetectable 15 days after bombardment. Expression of GUS activity wasrecorded mainly in somatic embryonal heads of various stages ofdevelopment and occasionally in suspensor cells. Similar to GUSexpression, modified green fluorescent protein (GFP) was detected in theembryonal head cells 24 hours after bombardment. GFP-expressingsuspensor cells were both more infrequent and difficult to detect, astheir highly vacuolate nature rendered the GFP presence less visibleagainst the yellow background autofluorescence.     

Studies on experimental chlamydial mastitis in goat histoenzymology

Two strains of Chlamydia psittaci (one isolated from aborted goat foetus and the other from brain of a buffalo calf that had died of meningoencephalitis) were injected intracisternally into six goats to produce experimental mastitis. Cryostat sections of 7–8 μm thickness, obtained from udder, teat, liver and kidney of infected and control animals were incubated for histoenzymic demonstration of alkaline-(AKPase), acid-(ACPase) and adenosine-tri-(ATPase) phosphatases; lactate-(LDH) and succinate-(SDH) dehydrogenases and for reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-D). Results demonstrated that AKPase and NADPH-D declined while ACPase accumulated in acinar cells of udder while both NADPH-D and ACPase decreased in teat sinus epithelium. Hepatic canaliculi in perilobular areas of liver lobules registered complete absence of AKPase and ATPase. Hepatocytes and renal tubules accumulated LDH, SDH and NADPH-D. The interstitial connective tissue of udder and kidney presented higher levels of AKPase. Comparison of results with biochemical alterations in the level of these enzymes revealed striking discrepancies which seem to arise because of failure of biochemical procedures to discriminate between functional cells of tissue and inflammatory cells. The functional significance of histoenzymic alterations has been discussed.