Qualitative risk assessment of transmission pathways of highly pathogenic avian influenza (HPAI) virus at live poultry markets in Dhaka city,Bangladesh

Analysis of environmental samples obtained from the Live Poultry Markets (LPMs) of Dhaka City, Bangladesh, has revealed that the highest degree of prevalence of highly pathogenic avian influenza A (HPAI, H5N1), besides other subtypes of the LPAI virus, poses the plausible risk of transmission of these viruses between human and poultry species. The present study was conducted using the OIE risk analysis framework to assess the risk level of each pathway successively. The estimated risk parameters were integrated towards to obtain the overall risk level for each specific HPAI transmission pathway using the matrix adapted by Cristobel Zepeda accompanying other expert consultations. The relevant data obtained from published and unpublished sources, together with survey data of field observations, were used to formulate and confirm the risk pathways and their associated risks. The results revealed that the risk of the release of the HPAI virus was medium when exposure was high. Additionally, the consequence would be considered very high with a medium degree of uncertainty for all parameters. Ultimately, the overall risk for transmission was estimated as medium with a medium degree of uncertainty. The findings of this study reveal that there is a significant threat that HPAI virus transmission could occur among poultry and humans and effectively sustain within the environment of the LPMs. Our findings are primarily focused on public health considerations, the hygienic slaughter of poultry and the relevant cleaning and sanitation practices conducted in the LPMs to support evidence‐based decision‐making processes. The findings of the study have the potential to be used to formulate effective risk reduction measures and can be further adapted in low‐resource settings without major infrastructural changes required of the LPMs. All of which would reduce the risk of HPAI virus release and further lessen the degree of exposure and transmission in established LPMs.     

Milk production of Jersey and Fleckvieh × Jersey cows in a pasture-based feeding system

Milk production parameters of purebred Jersey (J) cows and Fleckvieh?×?Jersey (F?×?J) cows in a pasture-based feeding system were compared using standard milk recording procedures. Milk, fat and protein production was adjusted to 305 days per lactation and corrected for age at calving. Effects of breed, parity, month and year were estimated for milk, fat and protein yield as well as fat and protein percentage, using the general linear model procedure. Fixed effects identified as affecting milk production parameters significantly were breed, parity and year. F?×?J cows produced significantly more milk than J cows (6141?±?102 and 5398?±?95 kg milk, respectively). Similarly, fat and protein yields were significantly higher in F?×?J (272?±?4 and 201?±?3 kg, respectively) than in Jersey cows (246?±?3 and 194?±?2 kg, respectively). Fat and protein percentages only differed slightly in absolute terms being 4.61?±?0.04 % fat in the Jersey compared to 4.47?±?0.04 % fat in the F?×?J. Protein levels for J and F?×?J cows were 3.62?±?0.03 and 3.51?±?0.03 %, respectively. Despite a lower fat percentage, F?×?J crossbred cows may be more productive than purebred Jersey cows which may be due to heterotic effects.     

Actin Polymerization: An Event Regulated by Tyrosine Phosphorylation During Buffalo Sperm Capacitation

In the female reproductive tract, the spermatozoa undergo a series of physiological and biochemical changes, prior to gaining the ability to fertilize, that result to capacitation. However, the actin polymerization and protein tyrosine phosphorylation are the two necessary steps for capacitation. In this study, we have demonstrated the actin polymerization and established the correlation between protein tyrosine phosphorylation and actin reorganization during in vitro capacitation in buffalo (Bubalus bubalis) spermatozoa. Indirect immunofluorescence and Western blot techniques were used to detect actin polymerization and tyrosine phosphorylation. The time‐dependent fluorimetric studies revealed that the actin polymerization starts from the tail region and progressed towards the head region of spermatozoa during capacitation. The lysophosphatidyl choline (LPC)‐induced acrosome reaction (AR) stimulated quick actin depolymerization. The inhibitor cytochalasin D (CD) blocked the in vitro capacitation by inhibiting the actin polymerization. In addition, we also performed different inhibitor (Genistein, H‐89, PD9809 and GF‐109) and enhancer (dbcAMP, H₂O₂ and vanadate) studies on actin tyrosine phosphorylation and actin polymerization. The inhibitors of tyrosine phosphorylation inhibit actin tyrosine phosphorylation and polymerization, whereas enhancers of tyrosine phosphorylation stimulate F‐actin formation and tyrosine phosphorylation. These observations suggest that the tyrosine phosphorylation regulates the actin polymerization, and both are coupled processes during capacitation of buffalo spermatozoa.     

Assessment of Disturbances in Growth and Physiology of Carrot Caused by Chromium Stress

To see the deleterious effects of excess chromium (Cr) on carrot (Daucus carota L.) the cv. ‘Pusa Keshari’ was grown in refined sand under controlled glasshouse conditions with a complete nutrient solution (without Cr) for 48 days. On the 49th day, pots with two plants each were separated into three sets. One set served as a control. In each of the other two sets, Cr was supplied as dichromate at 0.1 and 0.5 mM to the basal nutrient solution. At 0.5 mM Cr toxicity symptoms appeared at d 52 (4 days after Cr supply) as reduction in growth and leaf size and loss of turgor. Old leaves became chlorotic and wilted. Chlorosis intensified and turned necrotic in another few days. These symptoms spread to next upper leaf. The development of chlorosis in leaves was delayed in plants receiving 0.1 mM Cr. At excess Cr (0.5 mM) in carrot, the biomass, concentration of chlorophylls a and b, iron (Fe), sulfur (S), and phosphorus (P) in shoots, and activity of catalase (CAT) in leaves decreased whereas the concentration of Cr and the activity of peroxidase (POX), superoxide dismutase (SOD), ribonuclease (RNAse), and acid phosphatase (A P) in leaves increased.     

Role of crop diversification and integrated nutrient management in resilience of soil fertility under rice-wheat cropping system

A field study conducted for two crop cycles of five cropping systems supplied with six nutrient combinations at the Indian Agricultural Research Institute, New Delhi indicated that the cropping systems having a legume increased organic C content over initial level by 0.02?–?0.05%, available N by 3.5?–?14.1?kg ha^???1, whereas the rice-wheat cropping system resulted in a reduction in organic C and available N over initial level by 0.05% and 1.5?kg ha^???1, respectively after 2 years of study. Rice-potato-mungbean cropping system resulted in a negative balance of available P and rice-clover cropping system had a negative balance of both available P and available K content in soil and thus call for adequate P and K fertilization. Application of P and K helped in building up their content in soil; NPK?+?FYM showed the highest increase in organic C, available N, available P and available K content in soil. These results suggest the inclusion of a legume in a cropping system for maintaining organic C and available N in soil and adequate P and K fertilization for arresting the depletion of available P and K content in soil. Integrated nutrient management is one of the best methods for resilience of soil fertility under rice-wheat cropping system.     

Cryopreservation of Zona‐Free Cloned Buffalo (Bubalus Bubalis) Embryos: Slow Freezing vs Open‐Pulled Straw Vitrification

This study was carried out to compare the post‐thaw cryosurvival rate and the level of apoptosis in vitro produced zona‐free cloned buffalo blastocysts subjected to slow freezing or vitrification in open‐pulled straws (OPS). Zona‐free cloned embryos produced by handmade cloning were divided into two groups and were cryopreserved either by slow freezing or by vitrification in OPS. Cryosurvival of blastocysts was determined by their re‐expansion rate following post‐thaw culture for 22–24 h. The post‐thaw re‐expansion rate was significantly (p < 0.05) higher following vitrification in OPS (71.2 ± 2.3%) compared with that after slow freezing (41.6 ± 4.8%). For examining embryo quality, the level of apoptosis in day 8 frozen‐thawed blastocysts was determined by TUNEL staining. The total cell number was not significantly different among the control non‐cryopreserved cloned embryos (422.6 ± 67.8) and those cryopreserved by slow freezing (376.4 ± 29.3) or vitrification in OPS (422.8 ± 36.2). However, the apoptotic index, which was similar for embryos subjected to slow freezing (14.8 ± 2.0) or OPS vitrification (13.3 ± 1.8), was significantly (p < 0.05) higher than that for the control non‐cryopreserved cloned embryos (3.4 ± 0.6). In conclusion, the results of this study demonstrate that vitrification in OPS is better than slow freezing for the cryopreservation of zona‐free cloned buffalo blastocysts because it offers a much higher cryosurvival rate.     

Effect of Physiologically Relevant Heat Shock on Development,Apoptosis and Expression of Some Genes in Buffalo (Bubalus bubalis) Embryos Produced In Vitro

For investigating the effects of physiologically relevant heat shock, buffalo oocytes/embryos were cultured at 38.5°C (control) or were exposed to 39.5°C (Group II) or 40.5°C (Group III) for 2 h once every day throughout in vitro maturation (IVM), fertilization (IVF) and culture (IVC). Percentage of oocytes that developed to 8‐cell, 16‐cell or blastocyst stage was lower (p < 0.05) and the number of apoptotic nuclei was higher (p < 0.05) for Group III > Group II > controls. At both 8–16‐cell and blastocyst stages, relative mRNA abundance of stress‐related genes HSP 70.1 and HSP 70.2 and pro‐apoptotic genes CASPASE‐3, BID and BAX was higher (p < 0.05) in Groups III and II than that in controls with the exception of stress‐related gene HSF1. Expression level of anti‐apoptotic genes BCL‐XL and MCL‐1 was also higher (p < 0.05) in Groups III and II than that in controls at both 8–16‐cell and blastocyst stages. Among the genes related to embryonic development, at 8–16‐cell stage, the expression level of GDF9 was higher (p < 0.05) in Group III than that in controls, whereas that of GLUT1, ZAR1 and BMP15 was not significantly different among the three groups. At the blastocyst stage, relative mRNA abundance of GLUT1 and GDF9 was higher (p < 0.05) in Group II than that in controls, whereas that of ZAR‐1 and BMP15 was not affected. The results of this study demonstrate that exposure of buffalo oocytes and embryos to elevated temperatures for duration of time that is physiologically relevant severely compromises their developmental competence, increases apoptosis and affects stress‐, apoptosis‐ and development‐related genes.     

Buffalo (Bubalus bubalis) ES Cell–Like Cells are Capable of In Vitro Skeletal Myogenic Differentiation

When buffalo embryonic stem (ES) cell–like cells that expressed surface markers SSEA‐4, TRA‐1‐60, TRA‐1‐81, CD9 and CD90 and intracellular markers OCT4, SOX2 and FOXD3, as shown by immunofluorescence, and that expressed REX‐1 and NUCLEOSTEMIN as confirmed by RT‐PCR, were subjected to suspension culture in hanging drops in absence of LIF and buffalo foetal fibroblast feeder layer support, they differentiated to form three‐dimensional embryoid bodies (EBs). Of 231 EBs examined on Day 3 of suspension culture, 141 (61.3 ± 3.09%) were of compact type, whereas 90 (38.4 ± 3.12%) were of cystic type. The cells obtained from EBs were found to express NF‐68 and NESTIN (ectodermal lineage), BMP‐4 and α‐skeletal actin (mesodermal lineage), and α‐fetoprotein, GATA‐4 and HNF‐4 (endodermal lineage). When these EBs were cultured on gelatin‐coated dishes, they spontaneously differentiated to several cell types such as epithelial‐ and neuron‐like cells. When EBs were cultured in the presence of 1 or 2% DMSO or 10^?8 m or 10^?7 m retinoic acid for 25 days, ES cells could be directed to form muscle cell–like cells, the identity of which was confirmed by expression of α‐actinin by immunofluorescence and of MYF‐5, MYOD and MYOGENIN genes by RT‐PCR. MYOD was first detected on Day 10 in both treatment groups and on Day 15 in controls, whereas MYOGENIN was first detected on Day 10, Day 15 and Day 25 in the presence of retinoic acid, in the presence of DMSO and in controls, respectively. The present study demonstrates the ability of buffalo ES cell–like cells to undergo directed differentiation to cells of skeletal myogenic lineage.     

Nemafric-BL phytonematicide induces egg hatch inhibition in Meloidogyne incognita

Contribution of egg hatch inhibition in the overall reduction of Meloidogyne incognita numbers by Nemafric-BL phytonematicide was determined in vitro using a series of water-diluted phytonematicide solutions. Hatched juveniles were assessed at four different incubation periods. Thereafter, each phytonematicide solution was water-diluted 5 times and incubated for 5 days to evaluate the reversibility of egg hatch inhibition. At all exposure periods, except the shortest (24?h), concentration effects on egg hatch were highly significant with juvenile numbers versus increasing concentrations exhibiting quadratic relations. The models for the three consecutive exposure periods explain the quadratic relations by 95%, 94% and 98%, respectively. Results suggested that egg hatch inhibition was one of the mechanisms involved in reducing M. incognita population densities by Nemafric-BL phytonematicide.