Microclimate modification in Murrah buffalo (Bubalus bubalis) heifers during summer: Effect on intake, growth and hematobiochemistry.

The present investigation evaluated the effect of microclimate modification on feed intake, growth performance, and hemato-biochemical profile of Murrah buffalo (Bubalus bubalis) heifers during summer. Twenty-four buffalo heifers, between 15 and 20 months of age with an average body weight of 363.75 ± 11.27 kg, were randomly assigned to 4 groups based on their age and body weight. The heifers of the T0 (control) group were kept under the shed without any cooling treatment, while the animals in group T1 were tied with a cooling jacket. Buffalo heifers of group T2 were subjected to a cooling jacket with forced ventilation and animals in group T3 were treated with intermittent sprinkling (10 min., at 2 h intervals) and forced ventilation between 9.00 AM and 6.00 PM during the experiment. The ambient temperature inside the shed and core body temperature were reduced in groups T1, T2, and T3 compared to T0. Heifers had higher dry matter intake when subjected to cooling treatments T1, T2, and T3, whereas water intake was reduced in T2 and T3 groups. The animals in T2 and T3 groups attained higher average daily weight gain, while the feed conversion was better in the T3 group compared to T0. The hematological measures such as hemoglobin, total erythrocyte count, and total leucocyte count were found higher in T0. The serum glucose, sodium, and potassium levels increasedand alkaline phosphatase activity decreased in groups T1, T2 and T3 when compared with T0. It can be concluded that the provision of intermittent sprinkling and cooling jacket in combination with forced ventilation could improve the microclimate, which in turn could enhance the performance of Murrah heifers during hot summer days in the tropics.

Effect of milking environment enrichment through music on production performance and behaviour in cattle.

Enrichment of milking environment through music has been proposed to help animals to cope with divergent stressors. In sight of the above, a study was conducted to evaluate the effect of Indian instrumental music-based environmental enrichment played in yaman raga on milk production performance and behaviour in cattle. A total of 21 lactating dairy cattle (Vrindavani crossbred cows) having similar parity and stage of lactation were selected in three groups - T1, T2 and T3, each consisting of seven animals. The T1 and T2 groups were exposed to instrumental flute and sitar, respectively, 10 min prior to the start of milking and continued till completion of milking; while the T3 group served as control. Musical enrichment of the environment was done using recorded-tape of flute and sitar was played in yamen raga at 40-60 (dB) decibel intensity. The results revealed a non-significant difference in milk yield, rectal temperature, respiration rate, T3 (triiodothyronine) and T4 (thyroxine) hormones. However, there exhibited a significant (p < 0.05) difference in milking time, milking speed, cortisol hormones and behavioural parameters such as milk let-down in the animals exposed to music compared to the control group. Thus, the results have significant implications relating to the behavioural fitness and welfare of dairy animals and reducing residual milk.

Genetic parameters of pre-weaning weights in crossbred piglets using multi-trait animal model.

Variances and genetic parameters (heritability and correlation) were estimated for pre-weaning weights from birth to 6 weeks of age at weekly intervals in crossbred (Landrace X Desi) pigs. Data were analysed using multi-trait animal model incorporating dam's age (covariable), parity of sow, sex of piglet, season of birth and year of birth as fixed effects and direct additive genetic, maternal genetic and permanent litter as random effects. The estimates of direct additive genetic and maternal genetic heritability were 0.18 ± 0.04 and 0.14 ± 0.04; 0.73 ± 0.06 and 0.62 ± 0.06; 0.64 ± 0.06 and 0.37 ± 0.05; 0.42 ± 0.05 and 0.30 ± 0.05; 0.37 ± 0.04 and 0.33 ± 0.05; 0.43 ± 0.04 and 0.34 ± 0.06 and 0.47 ± 0.04 and 0.36 ± 0.05 for birth weight and subsequent weights at weekly intervals up to 6 weeks, respectively. The permanent litter effect ranged from 0.02 ± 0.01 (birth weight) to 0.11 ± 0.04 (weight at first week). The estimates of correlation between direct additive genetic component and maternal genetic component (i.e. ram) were moderate to high but all negative. ram ranged from - 0.79 ± 0.15 (weight at first week) to - 0.49 ± 0.26 (birth weight). The direct additive genetic and maternal genetic correlations between birth weight and weaning weight were 0.59 ± 0.04 and 0.62 ± 0.13, respectively, which revealed sufficient scope for selection based on birth weight itself. Early weights were found to be highly heritable with due influence of maternal and permanent litter effects, and thus, appropriate selection interventions may be aimed at this stage for favourable changes at subsequent ages.

The high frequency of non-aspartic acid residues at HA222 in influenza A(H1N1) 2009 pandemic viruses is associated with mortality during the upsurge of 2015: a molecular and epidemiological study from central India.

Influenza A(H1N1) viruses of the 2009 pandemic (A(H1N1)pdm09) continue to cause outbreaks in the post-pandemic period. During January to May 2015, an upsurge of influenza was recorded that resulted in high fatality in central India. Genetic lineage, mutations in the hemagglutinin (HA) gene and infection by quasi-species are reported to affect disease severity. The objective of this study is to present the molecular and epidemiological trends during the 2015 influenza outbreak in central India. All the referred samples were subjected to qRT-PCR for diagnosis. HA gene sequencing (23 survivors and 24 non-survivors) and cloning were performed and analyzed using Molecular Evolutionary Genomic Analyzer (MEGA 5·05). Of the 3625 tested samples, 1607 (44·3%) were positive for influenza A(H1N1)pdm09, of which 228 (14·2%) individuals succumbed to death. A significant trend was observed in positivity (P = 0·003) and mortality (P < 0·0001) with increasing age. The circulating A(H1N1)pdm09 virus was characterized as belonging to clade-6B. Clinically significant mutations were detected. Patients infected with the quasi-species of the virus had a greater risk of death (P = 0·009). This study proposes a robust molecular and clinical surveillance program for the detection and characterization of the virus, along with prompt treatment protocols to prevent outbreaks.

Maiden outbreaks of dengue virus 1 genotype III in rural central India.

Dengue is regarded as the most important arboviral disease. Although sporadic cases have been reported, serotypes responsible for outbreaks have not been identified from central India over the last 20 years. We investigated two outbreaks of febrile illness, in August and November 2012, from Korea district (Chhattisgarh) and Narsinghpur district (Madhya Pradesh), respectively. Fever and entomological surveys were conducted in the affected regions. Molecular and serological tests were conducted on collected serum samples. Dengue-specific amplicons were sequenced and phylogenetic analyses were performed. In Korea and Narsinghpur districts 37·3% and 59% of cases were positive, respectively, for dengue infection, with adults being the worst affected. RT-PCR confirmed dengue virus serotype 1 genotype III as the aetiology. Ninety-six percent of infections were primary. This is the first time that dengue virus 1 outbreaks have been documented from central India. Introduction of the virus into the population and a conducive mosquitogenic environment favouring increased vector density caused the outbreak. Timely diagnosis and strengthening vector control measures are essential to avoid future outbreaks.

Circulation of hepatitis A genotype IIIA virus in paediatric patients in central India.

Hepatitis A virus (HAV) infection, a major cause of childhood hepatitis is transmitted by orofaecal route. Children mostly suffer with subclinical infection but may have serious clinical implications leading to hospitalization and mortality. IgM ELISA and nRT PCR were conducted on the blood samples collected from HAV suspected paediatric cases referred to the viral diagnostic laboratory in the Regional Medical Research Centre for Tribals at Jabalpur, Central India. The nRT PCR products were sequenced and phylogenetic analysis was done. Of the 195 samples tested, 41 (21%) were positive for HAV antibodies, among which 38 (92%) belonged to paediatric age group and 32 per cent of these were hospitalized. nRT PCR and sequencing confirmed the presence of HAV. Phylogenic analysis revealed circulation of genotype III A in central India. Regular serological and molecular monitoring would aid in understanding epidemiology of HAV and plan intervention strategies.

First report of detection and molecular confirmation of Plasmodium ovale from severe malaria cases in central India.

OBJECTIVE: We report for the first-time detection of Plasmodium ovale in central India. METHODS: From 2010, all hospitalised suspected malaria cases at the malaria clinic of the Regional Medical Research Centre for Tribals in Bastar district, central India, were screened. Plasmodium species were identified by microscopy and species-specific nested PCR of 18s rRNA. RESULTS: Of 256 enrolled cases of confirmed P. falciparum malaria by microscopy, P. ovale infection was detected in three cases (1.2%) by PCR and sequencing. Of these three cases, one had cerebral malaria and another had severe malaria anaemia. In both of these cases, P. ovale infection was mixed with P. falciparum, while in third case the infection was mixed with both P. falciparum and P. vivax. Phylogenetic analysis revealed that these isolates showed closed homology with West African genotypes. CONCLUSION: All three hospitalised patients were originally residents of remote inaccessible forest villages and never moved out of their residence. This finding also has implications in malaria control and elimination as P. ovale causes relapses. This study highlights the need of molecular diagnosis of malaria species for appropriate treatment and control.

Detection of dengue virus 4 from central India.

BACKGROUND & OBJECTIVES: Dengue is an important arboviral disease. All four dengue virus serotypes are reported to be circulating in India. It is also known that different serotypes, genotypes and clades of genotype determine outbreak severity. Dengue affected children are known to have serious disease outcome. We carried out this study to give reliable diagnosis of dengue infection in children and to detect circulating serotype in central India. METHODS: Samples collected from paediatric patients suspected to have dengue fever were subjected to IgM and IgG ELISA to determine dengue virus infection. Samples collected within 0-5 days of onset of illness and positive by IgM ELISA were tested by nested reverse transcription polymerase chain reaction (nRT-PCR). The PCR products were sequenced and analyzed. RESULTS: Of the 89 samples tested, 18 and 7 were positive for dengue IgM and IgG, respectively. Dengue activity was observed in both Jabalpur city and adjoining rural settings. One sample found positive by nRT-PCR was further sequenced to confirm dengue virus 4 as aetiological agent. INTERPRETATION & CONCLUSIONS: Our findings demonstrated dengue virus infection in children and adolescent in central India. Because of continuous changing epidemiology, it is important to monitor dengue virus activity at both serological and molecular level in this part of the country for better patient care and management.

Mining and analysis of microsatellites in human coronavirus genomes using the in-house built Java pipeline.

Microsatellites or simple sequence repeats are motifs of 1 to 6 nucleotides in length present in both coding and non-coding regions of DNA. These are found widely distributed in the whole genome of prokaryotes, eukaryotes, bacteria, and viruses and are used as molecular markers in studying DNA variations, gene regulation, genetic diversity and evolutionary studies, etc. However, in vitro microsatellite identification proves to be time-consuming and expensive. Therefore, the present research has been focused on using an in-house built java pipeline to identify, analyse, design primers and find related statistics of perfect and compound microsatellites in the seven complete genome sequences of coronavirus, including the genome of coronavirus disease 2019, where the host is Homo sapiens. Based on search criteria among seven genomic sequences, it was revealed that the total number of perfect simple sequence repeats (SSRs) found to be in the range of 76 to 118 and compound SSRs from 01 to10, thus reflecting the low conversion of perfect simple sequence to compound repeats. Furthermore, the incidence of SSRs was insignificant but positively correlated with genome size (R2 = 0.45, p > 0.05), with simple sequence repeats relative abundance (R2 = 0.18, p > 0.05) and relative density (R2 = 0.23, p > 0.05). Dinucleotide repeats were the most abundant in the coding region of the genome, followed by tri, mono, and tetra. This comparative study would help us understand the evolutionary relationship, genetic diversity, and hypervariability in minimal time and cost.

Characterization of drug resistance and genetic diversity of Plasmodium falciparum parasites from Tripura, Northeast India.

Monitoring of anti-malarial drug resistance is vital in Northeast India as this region shares its international border with Southeast Asia. Genetic diversity of Plasmodium parasites regulates transmission dynamics, disease severity and vaccine efficacy. P. falciparum chloroquine resistance transporter (Pfcrt), multidrug resistance-1 (Pfmdr-1) and kelch 13 propeller (PfK-13) genes which govern antimalarial drug resistance and three genetic diversity markers, merozoite surface protein 1 and 2 (Pfmsp-1, Pfmsp-2) and glutamate rich protein (Pfglurp) were evaluated from Tripura, Northeast India using molecular tools. In the Pfcrt gene, 87% isolates showed triple mutations at codons M74I, N75E and K76T. 12.5% isolates in Pfmdr-1 gene showed mutation at N86Y. No polymorphism in PfK-13 propeller was found. Polyclonal infections were observed in 53.85% isolates and more commonly in adults (p = 0.0494). In the Pfmsp-1 locus, the K1 allelic family was predominant (71.2%) followed by the 3D7/IC family (69.2%) in the Pfmsp-2 locus. RII region of Pfglurp exhibited nine alleles with expected heterozygosity of 0.85. The multiplicity of infection for Pfmsp-1, Pfmsp-2 and Pfglurp were 1.56, 1.31 and 1.06 respectively. Overall, the study demonstrated a high level of chloroquine resistance and extensive parasite diversity in the region, necessitating regular surveillance in this population group.

Antifolate drug resistance: Novel mutations and haplotype distribution in dhps and dhfr from Northeast India.

Malaria is a major public health concern in Northeast India with a preponderance of drug-resistant strains. Until recently the partner drug for artemisinin combination therapy (ACT) was sulphadoxine pyrimethamine (SP). Antifolate drug resistance has been associated with the mutations at dihydropteroate synthase (dhps) and dihydrofolatereductase (dhfr) genes. This study investigated antifolate drug resistance at the molecular level. A total of 249 fever cases from Arunachal Pradesh, NE India, were screened for malaria, and of these, 75 were found to be positive for Plasmodium falciparum. Samples were sequenced and analysed with the help of BioEdit and ClustalW. Three novel point mutations were found in the dhps gene with 10 haplotypes along with the already reported mutations. A single haplotype having quadruple mutation was found in the dhfr gene. The study reports higher degree of antifolate drug resistance as evidenced by the presence of multiple point mutations in dhps and dhfr genes. The findings of this study strongly discourage the use SP as a partner drug in ACT.