Development of a novel sensitive single-tube nested PCR assay for the detection of African swine fever virus.

African swine fever (ASF) is a highly fatal and contagious viral disease caused by African swine fever virus (ASFV). It has caused significant economic losses to the swine industry and poses a serious threat to food security worldwide. Diagnostic tests with high sensitivity are essential for the effective management of ASF. Here, we describe a single-tube nested PCR (STN-PCR) assay for the detection of ASFV in which two consecutive amplification steps are carried out within a single tube. Two pairs of primers (outer and inner) were designed to target the p72 gene of ASFV. The primer concentrations, annealing temperatures, and number of amplification cycles were optimized to ensure the consecutive utilization of outer and inner primer pairs during amplification while minimizing the likelihood of amplicon contamination. In comparison with two conventional endpoint PCR assays (one of which is recommended by the World Organization for Animal Health), the newly developed STN-PCR assay demonstrated a 100-fold improvement in the limit of detection (LOD), detecting 100 copies of ASFV genomic DNA, whereas the endpoint PCR assays could detect no fewer than 10,000 copies. The clinical performance of the STN-PCR assay was validated using 95 tissue samples suspected of being positive for ASFV, and the assay showed 100% specificity. A Cohen's kappa value of 0.91 indicated perfect agreement between the assays. This new STN-PCR assay is a potentially valuable tool that will facilitate the control of ASF.

Effects of integrated nutrient management on sucrose phosphate synthase enzyme activity and grain quality traits in rice.

Cropping systems and management practices can greatly affect rice crop, finally determining their grain yield and quality. In this study, we evaluated the effects of integrated nutrient management practices on sucrose phosphate synthase enzyme (SPS) activity and grain quality traits of rice. Field experiments were conducted at North Bank Plain Agro-climatic Zone of Assam, India from 2013 to 2015 on three rice ecosystems (winter, summer, and pre-monsoon). Selected fertilizer treatments were NPK as recommended inorganic fertilizer, NPK + cow dung, NPK + whole parts of the green manure Sesbania aculeata, NPK + compost of Azolla caroliniana and NPK + rice husk dust. NPK + Azolla compost application resulted in higher SPS activity compared to control. A significant relationship between panicle biomass and SPS enzyme activity was observed in the rice plants grown in different ecosystems. Integrated nutrient fertilizers in rice soil had a significant impact on the grain quality of rice. Grain nitrogen and crude protein content were higher at NPK + green manure Sesbania aculeata applied field irrespective of the ecosystems. NPK and Azolla compost were effective in improving grain productivity and grain quality parameters viz; total carbohydrates, starch, and amylose in rice crop. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01148-w.

Tape strips in dermatology research.

Tape strips have been used widely in dermatology research as a minimally invasive method to sample the epidermis, avoiding the need for skin biopsies. Most research has focused on epidermal pathology, such as atopic eczema, but there is increasing research into the use of tape strips in other dermatoses, such as skin cancer, and the microbiome. This review summarizes the technique of tape stripping, and discusses which dermatoses have been studied by tape stripping and alternative minimally invasive sampling methods. We review the number of tape strips needed from each patient and the components of the epidermis that can be obtained by tape stripping. With a focus on protein and RNA extraction, we address the techniques used to process tape strips. There is no optimal protocol to extract protein, as this depends on the abundance of the protein studied, its level of expression in the epidermis and its solubility. Many variables can alter the amount of protein obtained from tape strips, which must be standardized to ensure consistency between samples. No study has compared different RNA extraction techniques, but our own experience is that RNA yield is optimized by using 20 tape strips and the use of a cell scraper.

Productive and reproductive performances of two-breed and three-breed pig crosses with Niang Megha, Hampshire and Duroc inheritance reared under subtropical Eastern Himalayan hilly climate.

The objective of the study was to evaluate the performance of different crossbreeds, viz., two-breed crosses including HN-50 (50% Hampshire × 50% Niang Megha) and HN-75 (75% Hampshire × 25% Niang Megha) and three-breed cross, HND (25% Hampshire × 25% Niang Megha × 50% Duroc) for suggesting suitable crossbred pigs with appropriate inheritance for subtropical Eastern Himalayan hilly climate. These crossbreed pigs were reared in standard management conditions in Nucleus Pig Breeding Farm of ICAR RC for NEH region. A total of 1995 records were collected which included data on production performance (n = 1466), reproductive performance (n = 428) and carcass traits (n = 101) from farm record book maintained over a period of 7 years. Productive performance included body weight, ADG and FCR at different stages of growth. The study revealed productive performance was highest in two-breed cross of pigs with 75% H inheritance (HN-75) and three-breed cross (HND) pigs. Reproductive performance included ages at puberty, first conception and first farrowing, along with inter-farrowing interval, pregnancy and farrowing rate as well as litter performance. The HN-75 was found to be having shorter inter-farrowing interval and higher pregnancy rate than other genetic groups. Regarding carcass traits, three-breed cross had a higher dressing percentage and less back-fat thickness than other crossbred pigs. Two breed crosses of pigs were found to be having a higher back-fat thickness than three-breed cross pig, and HN-75 had a better dressing percentage than HN-50. Thus, it was concluded that three-breed cross was recommended for lean meat production, and two-breed cross HN-75 was recommended for both breeding and fattening purposes for subtropical Eastern Himalayan hilly climate.

Characterization of phenotype variations of luminescent and non-luminescent variants of Vibrio harveyi wild type and quorum sensing mutants.

Vibrio harveyi, a luminescent Gram-negative motile marine bacterium, is an important pathogen responsible for causing severe diseases in shrimp, finfish and molluscs leading to severe economic losses. Non-luminescent V. harveyi obtained by culturing luminescent strains under static and dark condition were reported to alter the levels of virulence factors and metalloprotease gene and luxR expression when compared to their luminescent variants. Presently, we conducted an in vitro study aiming at the characterization of virulence-related phenotypic traits of the wild-type V. harveyi BB120 strain and its isogenic quorum sensing mutants before and after switching to the non-luminescent status. We measured the production of caseinase, haemolysin and elastase and examined swimming motility and biofilm formation. Our results showed that switching from the bioluminescent to the non-luminescent state changed the phenotypic physiology or behaviour of V. harveyi resulting in alterations in caseinase and haemolytic activities, swimming motility and biofilm formation. The switching capacity was to a large extent independent from the quorum sensing status, in that quorum sensing mutants were equally capable of making the phenotypic switch.

Effects of low-density lipoproteins as additive on quality parameters and oxidative stress following cryopreservation of mithun (Bos frontalis) spermatozoa.

Artificial breeding of mithun poses several challenges including lack of standard protocol for cryopreservation of spermatozoa. This is further complicated by harmful effects of hen's egg yolk (EY) as additive in extender. Purified low-density lipoproteins (LDL) extracted from EY have been shown as beneficial over EY extender for long-term semen storage in several species. This investigation explored use of LDL versus EY on semen quality and oxidative stress following freezing-thawing of spermatozoa. A total of 25 of 50 ejaculates based on biophysical parameters were selected for the experiment. After diluting with the Tris-citrate-glycerol (TCG) extender, each sample was split into three equal aliquots: Group I, control, EY; Group II and Group III contained 8% and 10% purified LDL, respectively. Frozen-thawed samples were evaluated for motility parameters (progressive, and in the bovine cervical mucus penetration test [BCMPT]), viability, sperm and nuclear abnormality, acrosome integrity, and enzymatic (leakage of intracellular contents) and biochemical (oxidative stress) profiles and in vitro fertility (IVF) assay. Study revealed a significant (p < .05) improvement in viability, sperm and nuclear abnormality, acrosome integrity, motility (progressive and in cervical mucus), cholesterol content, and reduction in the leakage of intracellular enzymes in Group II. Moreover, intactness of acrosome and biochemical membranes was protected significantly (p < .05) in addition to significant (p < .05) improvement in binding per cent and binding index in IVF assay in extender containing 8% LDL. These results demonstrate that although cryopreservation of mithun's spermatozoa in EY was comparable with other species, addition of 8% LDL holds a clear advantage over EY or 10% LDL.

Emergence of dengue in tribal villages of Mandla district, Madhya Pradesh, India.

BACKGROUND & OBJECTIVES: Dengue (DEN) is a rapidly spreading arboviral disease transmitted by Aedes mosquitoes. Although it is endemic in India, dengue virus (DENV) infection has not been reported from tribal areas of Madhya Pradesh. Investigations were conducted to establish the aetiology of sudden upsurge of cases with febrile illness in June 2013 from tribal villages of Mandla district of Madhya Pradesh, India. METHODS: The rapid response team of the National Institute for Research in Tribal Health, Jabalpur, conducted clinical investigations and field surveys to collect the samples from suspected cases. Samples were tested using molecular and serological tools. Collected mosquitoes were identified and tested for the presence of virus using semi nested reverse transcriptase-polymerase chain reaction (nRT-PCR). The sequences were analysed to identify serotype and genotype of the virus. RESULTS: Of the 648 samples collected from 18 villages of Mandla, 321 (49.53%) were found to be positive for dengue. The nRT-PCR and sequencing confirmed the aetiology as dengue virus type 2. Eighteen per cent of patients needed hospitalization and five deaths were attributed to dengue. The virus was also detected from Aedes aegypti mosquito, which was incriminated as a vector. Phylogenetic analysis revealed that the dengue virus 2 detected belonged to cosmopolitan genotype of the virus. INTERPRETATION & CONCLUSIONS: Dengue virus serotype 2 was detected as the aetiological agent in the outbreak in tribal villages of Mandla district of Madhya Pradesh. Conducive man-made environment favouring mosquitogenic conditions and seeding of virus could be the probable reasons for this outbreak. Urgent attention is needed to control this new threat to tribal population, which is already overburdened with other vector borne diseases.

Effect of concentration and addition method of glycerol on the quality of cryopreserved mithun (Bos frontalis) spermatozoa.

The effect of concentration and addition method of glycerol on the quality of cryopreserved mithun (Bos frontalis) spermatozoa was investigated. Semen samples were collected from five healthy mithun bulls through rectal massage method and cryopreserved in liquid nitrogen. The samples were diluted in Tris-egg yolk-glycerol extender, equilibrated for 4 h at 4 °C and loaded into 0.50-ml straws. The straws were then frozen in liquid nitrogen vapour for 10 min and finally plunged into liquid nitrogen for storage. The required amount of glycerol was added into the diluted samples either in a single dose (3%, 4%, 5%, 6% or 7%; added at 37 °C immediately before equilibration) or in split doses (5%, 6% or 7%; the total amount was divided into four equal parts, and a part was added at 37 °C immediately before equilibration, and the remaining parts were added subsequently at 1, 2 and 3 h of equilibration at 4 °C). In the single-dose addition method, following freeze-thawing, greater (p < 0.05) motility (%) and proportion of live spermatozoa with intact acrosome (LSIA, %) in 5% glycerol (40.6 ± 1.7 and 43.4 ± 1.8 respectively) and lesser (p < 0.05) total morphological abnormalities (%) in 5% (14.1 ± 0.8) and 6% (13.7 ± 1.0) glycerol were observed compared to the other glycerol concentrations. In the split-dose addition method, following freeze-thawing, greater (p < 0.05) motility (%) and LSIA proportion (%) were found in 5% (50.2 ± 1.9 and 53.3 ± 1.8 respectively) compared to 6% or 7% glycerol, but the total morphological abnormalities were not different among the glycerol concentrations. In addition, in all the glycerol concentrations, better (p < 0.05) post-freeze-thaw motility and LSIA proportions were observed when glycerol was added in split doses compared to a single dose. In conclusion, Tris-egg yolk extender with 5% glycerol added in split doses was found most suitable for cryopreserving mithun sperm.

Zika virus in India: past, present and future.

Zika virus (ZIKV) is an arthropod-borne flavivirus that presents with acute febrile illness associated with rash, arthralgia and conjunctivitis. After years of sporadic reports in Africa, the three major outbreaks of this disease occurred in Yap Islands (2007), French Polynesia (2013-14) and South Americas (2015-16). Although, serological surveys suggested the presence of ZIKV in India in 1950s, cross-reactivity could not be ruled out. The first four proven cases of ZIKV from India were reported in 2017. This was followed by major outbreaks in the states of Rajasthan and Madhya Pradesh in 2018. Fortunately, the outbreaks in India were not associated with neurological complications. These outbreaks in India highlighted the spread of this disease beyond geographical barriers owing to the growing globalization, increased travel and ubiquitous presence of its vector, the Aedes mosquito. In this review, we discuss the epidemiology, clinical features and management of ZIKV in India.

Development of a novel visual isothermal amplification assay for rapid detection of Brucella spp.

Brucellosis is an economically important livestock disease worldwide besides having a noteworthy impact on human health. In this study, a rapid, simple, and ultra-sensitive nuclei-acid diagnostic technique was developed for the detection of brucellosis harnessing saltatory rolling circle amplification (SRCA). The diagnostic method was developed using World Organization for Animal Health (WOAH) approved primers targeting the bcsp31 gene of the Brucella genome. The assay can be accomplished within 90 min at a temperature of 65 °C without the requirement of sophisticated instrumentation. The result interpretation can be done with the naked eye with the aid of SYBR green dye. The developed technique displayed 100% specificity by amplifying only 10 reference and field strains of Brucella spp. and there was no cross-reactivity with the other tested pathogens. The lower limit of detections of SRCA and end-point PCR assays were 9.7 fg/µL (2.7 genome copies of Brucella) and 970 fg/µL, respectively. Thus, the developed SRCA assay was found to be 100× more sensitive than the end-point PCR assay. To the best of our knowledge, our study is the first one to develop an SRCA-based assay for the detection of brucellosis and it can be a diagnostic tool for resource-constrained laboratories and veterinary hospitals.

Plant morphophysiological and anatomical factors associated with nitrous oxide flux from wheat (Triticum aestivum).

Experiments were conducted to study the dynamics of nitrous oxide (N2O) emission from wheat varieties viz., Sonalika, HUW 468, HUW 234 and DBW 14 grown in alluvial soils of North Bank Plain Agroclimatic Zone of Assam, India. Attempts were made to find out the relationship of N2O emission with plant morphophysiological, anatomical and soil properties. N2O fluxes from wheat varieties ranged from 40 µg N2O-N m⁻² h⁻¹ to 295 µg N2O-N m⁻² h⁻¹. Soil organic carbon and soil temperature have shown significant relationship with N2O flux. The rate of leaf transpiration recorded from the wheat varieties at different growth stages exhibited a positive correlation with N2O emission suggesting that movement of N2O along with the transpirational water flow may be an important mechanism of N2O transport and emission through wheat plants. Anatomical investigation by scanning electron microscope revealed that N2O emission has relationship with stomatal frequency of leaf and leaf sheaths. Variety HUW 234 with the highest stomatal frequency of leaf and leaf sheath also recorded higher seasonal N2O emission compared to other varieties. Seasonal N2O emission (E(sif)) of the varieties ranged from 3.25 to 3.81 kg N2O-N ha⁻¹. Significant variations in E(sif) values were recorded within the varieties.

Effect of method and time of first colostrum feeding on serum immunoglobulin concentration, health status and body weight gain in mithun (Bos frontalis) calves.

The effect of method and time of first colostrum feeding on the concentration of serum immunoglobulin (Ig) was evaluated in mithun (Bos frontalis) calves. The hypotheses were that the variable method and time of first colostrum feeding might affect the level of serum Ig and in turn the growth performance and health status of the claves during the early age. The newborn calves were randomly allotted to one of the four experimental groups - G-1: allowed to suckle the dam at own choice, G-2: separated immediately after birth and allowed to suckle the dam first at 6 h and then at own choice, G-3: bottle fed ad libitum colostrum of its own dam first at 6 h and then at 6-h intervals until 24 h, G-4: bottle fed ad libitum colostrum of its own dam within 1 h, at 6 h and then at 6-h intervals until 24 h. The concentrations of IgG, IgM, and IgA were lowest (p < 0.01) at birth and increased following colostrum feeding irrespective of the experimental group. Highest concentrations of all the Ig classes were observed at 12-24 h after birth. The concentrations then transiently decreased from day 7 to 14, and then steadily increased after day 28. The concentrations of IgG (p < 0.01) and IgA (p < 0.05) were higher in G-1 in relation to the other groups during the first week after birth. Similarly, higher concentration of IgA (p < 0.05) was found in G-1 in relation to the other groups during the rest of the experimental period. The apparent absorption efficiency of colostral IgG was higher (p < 0.05) in G-4 in relation to G-3. Growth rate and health status were not influenced by the method and time of first colostrum feeding. In conclusion, a 6-h delay in the first colostrum feeding reduced the level of serum Ig noticeably.

Intake, digestibility and nitrogen balance in Mithun (Bos frontalis) offered urea-treated paddy straw based feed blocks.

The aim of the study was to determine the effect of feeding feed blocks containing varying proportion of urea-treated paddy straw (UTPS) on dry matter (DM) intake (DMI), nutrient utilization and N balance in Mithun. For the purpose, four adult male Mithun (279.5 ± 8.2 kg) were selected and offered four experimental rations viz. R(1) (Napier fodder + concentrate at 60:40), R(2) (UTPS + concentrate at 50:50), R(3) (UTPS + concentrate at 60:40) and R(4) (UTPS + concentrate at 70:30) in 4 × 4 Latin square design. The DMI % of body weight was 2.59, 2.96, 2.85 and 2.77 and the DMI g kg(-1) W(0.75) was 107, 123, 118 and 115 in Mithun fed R(1), R(2), R(3) and R(4), respectively. The mean DMI was (P < 0.01) higher in animals fed R(2) and R(3) than R(1) and R(4), whereas the water intake was (P < 0.01) higher in Mithun fed R(2), R(3) and R(4) than R(1). The digestibility of DM, organic matter, crude protein, crude fibre, neutral detergent fibre and cellulose were (P < 0.05) higher in animals fed R(2), R(3) and R(4) than R(1). A positive N balance was observed in all the experimental animals, with higher (P < 0.05) values among the animals offered R(2), R(3) and R(4) than R(1). The digestible crude protein and total digestible nutrient intakes were higher (P < 0.05) in Mithun fed R(2) and R(3) than R(1) and R(4). It is concluded that the UTPS can be incorporated up to 70% to formulate the complete feed/feed block and can be used for feeding of Mithun under complete confinement system.

Comparative nutrient utilization, growth, and rumen enzyme profile of mithun (Bos frontalis) and Tho-tho cattle (Bos indicus) fed on tree-leaves-based ration.

A study was conducted to compare the nutrient utilization, growth, and rumen enzyme profile of mithun (Bos frontalis) and Tho-tho cattle (Bos indicus) reared in the same feeding and managemental conditions. For the purpose, male mithun (n = 8) and male Tho-tho cattle (n = 8) of 1.5 years age, selected from the farm of National Research Centre on Mithun, Nagaland, India, were fed on mixed-tree-leaves-based ration as per the requirement of NRC (2001) for cattle for 12 months. Average daily gain (ADG), average dry matter intake (DMI), and feed conversion ratio (FCR) for all animals were recorded. A metabolic trial was conducted at 6 months of the experiment to assess the digestibility coefficient of different nutrients and nutritive value of ration. At 12 months of the experiment, rumen liquor was collected from all animals and analyzed for rumen enzyme profiles, viz., carboxymethylcellulase, xylanase, α-amylase, ß-glucosidase, α-glucosidase, urease, and protease. It was found that ADG (507.8 g vs 392.8 g), DM intake (6.59 vs 5.85 kg/day) and DMI/W(0.75) (98.75 g vs 91.00 g/day), crude protein intake (780 vs 700 g/day), and total digestible nutrient intake (3.65 vs 3.32 kg/day) were higher (p < 0.05) in mithun than cattle. The nitrogen balance was higher and FCR was better (p < 0.05) in mithun compared with cattle. The digestibility coefficient of different nutrients was similar (p > 0.05) between the species. The microbial enzyme profiles of mithun and cattle were not different (p > 0.05). The better growth performance of mithun than cattle as found in the present study clearly indicates that the mithun has higher genetic potential for growth than Tho-tho cattle of north-eastern hilly region of India.

Detection of bovine herpesvirus 1 sequences in yaks (Bos grunniens) with keratoconjunctivitis, using a highly sensitive nested polymerase chain reaction.

Thirty-seven yaks (Bos grunniens) with keratoconjunctivitis and 22 healthy yaks were used to investigate the role of bovine herpesvirus 1 (BoHV-1) in keratoconjunctivitis in yaks. Nucleic acid sequences of BoHV-1 glycoproteins B and E were detected in conjunctival swabs from all yaks with keratoconjunctivitis using a nested polymerase chain reaction (PCR). In 21 yaks, BoHV-1 sequences were detected along with Moraxella bovis (M. bovis) and Neisseria spp. The amplified BoHV-1 sequences were identical, and no nucleotide variation was observed when compared with a BoHV-1 reference strain using single-strand conformation polymorphism analysis of the amplified DNA sequences. Interestingly, BoHV-1 sequences could not be detected in samples from healthy yaks. However, conjunctival swabs from two healthy yaks (9.09%) yielded M. bovis and Neisseria spp. Samples from 35 yaks with keratoconjunctivitis showed positive reactions in an avidin biotin enzyme-linked immunosorbent assay for BoHV-1 antibodies; all the healthy yaks were seronegative. This is the first report of a possible association of BoHV-1 with keratoconjunctivitis in yaks.

Plant physiological and soil characteristics associated with methane and nitrous oxide emission from rice paddy.

Methane (CH4) and nitrous oxide (N2O) are important greenhouse gases causing global warming and climate change. Efforts were made to analyze the CH4 and N2O flux in relation to plant and soil factors from rice (Oryza sativa L.) paddy. Ten popularly grown rice varieties namely Rashmisali, Bogajoha, Basmuthi, Lalkalamdani, Choimora (traditional varieties); Mahsuri, Moniram, Kushal, Gitesh and Profulla (high yielding varieties = HYV) were grown during monsoon season of July 2006. The CH4 and N2O emissions were measured the date of transplanting onwards at weekly interval along with soil and plant parameters. The seasonal integrated CH4 and N2O emission (Esif) from rice ranged from 8.13 g m(-2) to 13.00 g m(-2) and 121.63 mg N2O-N m(-2) to 189.46 mg N2O-N m(-2), respectively. Variety Gitesh emitted less N2O and CH4 amongst all the rice varieties. Both CH4 and N2O emission exhibited a significant positive correlation with leaf area, leaf number, tiller number and root dry weight. Soil organic carbon of the experimental field was associated with both CH4 and N2O emission whereas nitrate-N content of soil was associated with N2O emission. Methane emission showed significant positive correlations with soil temperature and crop photosynthetic rate. Traditional rice varieties with profuse vegetative growth recorded higher CH4 and N2O fluxes compared to HYVs. Gitesh and Kushal having low seasonal CH4 and N2O emission with higher yield potential can be recommended as low greenhouse gas emitting rice varieties.

Slow release exogenous melatonin modulates scrotal circumference and testicular parameters, libido, endocrinological profiles and antioxidant and oxidative stress profiles in mithun.

Mithun (Bos frontalis) is a unique domestic free-range bovine species available in North Eastern hilly (NEH) regions of India and is reared for its protein rich meat. Although, mithun suffers from severe non-cyclical population fluctuations, it is not yet declared as endangered. Therefore, present study was conducted to assess the effect of slow release subcutaneous exogenous melatonin (MT) implant on sexual behaviour, scrotal circumference (SC) and testicular parameters, endocrinological profiles and antioxidant and oxidative stress profiles in mithun during different seasons (winter, spring, summer and autumn) to improve its reproductive efficiency. Experimental mithun bulls (5-6 years of age) were selected and divided randomly into two groups, Gr I: Control (n = 6) and Gr II: Treatment (n = 6; melatonin implant @ 18mg/50 kg B. Wt). Scrotal circumference and testicular parameters [total testicular volume and weight], endocrinological profiles [follicle stimulating hormone (FSH), luteinizing hormone/interstitial cell stimulating hormone (LH/ICSH), testosterone, thyroxine (T4), cortisol, prolactin and melatonin (MT)], sex behaviour profiles [libido score, mating ability score and sexual behaviour score], antioxidant profiles [total antioxidant capacity (TAC), catalase (CAT), glutathione (GSH), glutathione reductase (GSHRx) and superoxide dismutase (SOD)] and oxidative stress profile [malondialdehyde (MDA)] were estimated. Statistical analysis revealed that these experimental profiles differed between treatment and control groups within the seasons and among the seasons within the experimental groups. FSH, LH, testosterone, T4 and MT were higher and cortisol and prolactin were lower in spring and winter than in summer season. Concentration of FSH, LH, testosterone and MT were higher and concentration of T4, cortisol and prolactin were lower in MT treated than in untreated control group. Similarly, sexual behavioural scores, SC, testicular parameters and antioxidant profiles were higher and oxidative stress profile was lower in MT treated than in untreated control group and in winter and spring than in summer season. The study concluded that exogenous slow-release melatonin implantation and spring and winter seasons had significant beneficial effects in improvement of the antioxidant profiles, minimization of oxidative stress with cascading beneficial effects on endocrinological profiles, SC, testicular parameters and libido, which will improve the semen production, higher sperm cryo-survivability and fertility rate in mithun species.

Dietary high protein and vitamin C mitigates endosulfan toxicity in the spotted murrel, Channa punctatus (Bloch, 1793).

The amelioration effect of dietary high protein and vitamin C against stress was evaluated in spotted murrel, Channa punctatus, exposed to endosulfan. Two hundred and forty fish (average weight: 27.01 g/fish), distributed equally into 4 different groups (control, T(1), T(2), and T(3)), each with 6 replicates were fed with control (40% crude protein, CP and 0.1% vitamin C), T(1) (40% CP and 0.1% vitamin C), T(2) (50% CP and 0.1% vitamin C), and T(3) (50% CP and 0.2% vitamin C) diets for 90 days. Groups fed T(1), T(2), and T(3) diets were exposed to sublethal endosulfan concentration, whereas the control was maintained without endosulfan exposure. Results indicated significant reduction in the growth performance, survival, and activities of lactate dehydrogenase (liver and muscle), malate dehydrogenase (liver and muscle), enzymes of protein metabolism (aspartate amino transferase in liver and alanine amino transferase in liver and muscle), acetyl choline esterase (brain), alkaline phosphatase activity (liver), and ATPase (gill) enzymes of group fed control diet and exposed to endosulfan. However, endosulfan exposed fish fed high CP and vitamin C diet exhibited significant (P<0.05) improvement in their growth performance and metabolic enzyme activities. Further, high CP and vitamin C diet reduced endosulfan accumulation in the muscle. Overall results indicate that vitamin C (0.2%) supplementation in high CP (50%) diet improves growth, metabolism, and reduce endosulfan bioaccumulation in C. punctatus.

Biochemical and stress responses of rohu Labeo rohita and mrigal Cirrhinus mrigala in relation to acclimation temperatures.

The biochemical and stress responses of two Indian major carps, rohu Labeo rohita and mrigal Cirrhinus mrigala were studied after acclimating them to four preset temperatures (26, 31, 33 and 36 degrees C) for 30 days. The blood glucose and liver glycogen levels showed an inverse trend in both the species and were significantly different in L. rohita at higher temperatures. The decrease in the liver glycogen level of C. mrigala, however, was not significant. Plasma cortisol levels increased significantly whereas the ascorbic acid content in the brain and kidney of both the species decreased significantly with increasing temperatures. Total lipid content in the liver of both the species decreased significantly with increasing acclimation temperatures. The phospholipid concentration decreased in L. rohita with increasing acclimation temperatures, and in C. mrigala the values decreased up to 33 degrees C and increased at 36 degrees C. In C. mrigala, the cholesterol level decreased up to 33 degrees C and then increased at 36 degrees C, but the absolute value was lower in comparison to L. rohita. The cholesterol levels, however, were not significantly different in L. rohita. Triglycerides and free fatty acids concentrations decreased significantly with increasing acclimation temperatures in both the species. The present study indicates species-specific metabolic responses of L. rohita and C. mrigala to thermal acclimation.

Adaptive Radiotherapy for Carcinoma of the Urinary Bladder: Long-term Outcomes With Dose Escalation.

AIMS: To report long-term outcomes with dose-escalated, image-guided adaptive radiotherapy (ART) for bladder preservation in muscle-invasive bladder cancer (MIBC). MATERIALS AND METHODS: All MIBC patients receiving bladder-preserving ART at our institute from 2009 to 2018 were analysed. For ART, three anisotropic planning target volumes (PTV) were concentrically grown around the simulation bladder volume. A library of intensity-modulated radiotherapy plans was created for each patient. A total dose of 64 Gy in 32 fractions to the entire bladder and 55 Gy to pelvic nodes was planned, with 68 Gy to the tumour bed (2 Gy equivalent dose = 68.7 Gy, α/ß = 10) as simultaneous integrated boost for solitary tumours. The most appropriate PTV encompassing the bladder ('plan-of-the-day') was chosen daily using on-board megavoltage imaging. Neoadjuvant and concurrent chemotherapy was prescribed for medically fit patients. RESULTS: Of a total of 106 patients, most had T2 (68%) or T3 (19%) disease. Ninety-two patients (87%) completed 64 Gy to the whole bladder. Sixty-three patients (59%) received 68 Gy as tumour bed boost. Seventy-six per cent received concurrent weekly chemotherapy. At a median follow-up of 26 months, 3-year locoregional control, disease-free survival and overall survival were 74.3, 62.9 and 67.7%, respectively. Eighty-two per cent of patients retained disease-free bladder. Radiation Therapy Oncology Group grade III/IV acute genitourinary and gastrointestinal toxicities were 7.5% and 0%, respectively, and late genitourinary/gastrointestinal toxicities were 6.5% and 3.8%, respectively. Overall survival, disease-free survival, locoregional control and grade III/IV genitourinary/gastrointestinal toxicities did not differ significantly with dose escalation. CONCLUSION: Plan-of-the-day ART is clinically safe and effective for bladder preservation and can be implemented in routine clinical practice. A high bladder preservation rate is achievable without compromising on survival or toxicities. Dose escalation does not seem to affect outcomes.