Differential expression of inducible nitric oxide synthase and cytokine mRNA in chicken lines divergent for cutaneous hypersensitivity response

Phytohemagglutinin (PHA)-induced delayed-type hypersensitivity is an immunocompetent trait considered an indicator of cell-mediated immune or T-cell responses. Divergent selection was performed to generate high and low lines for response to PHA-P. Extreme-responder birds of the F2 generation in each line were used to study possible differences in macrophage activity and the associated functional genes. To evaluate macrophage activity, nitric oxide (NO) was estimated both systemically in serum and in in vitro monocyte culture. Semi-quantitative RT-PCR was used to detect the differential mRNA expression patterns of iNOS and MIP-1beta in monocyte culture, whereas T(H)1 cytokines (IL-2 and IFN-gamma) were studied in peripheral blood mononuclear cells (PBMC) at different time intervals after lipopolysaccharide (LPS) induction. The high line showed strong systemic, as well as in vitro NO production, compared to the low line, upon stimulation with NDV and LPS, similar to early and high iNOS mRNA expression. Following the pattern of iNOS gene expression, an early strong expression of cytokines with powerful iNOS-inducing action, such as IFN-gamma and the chemokine MIP-1beta, was observed in the high line. In contrast, for response to PHA-P, low expression of IL-2 was observed in the high compared to the low line. In conclusion, the study revealed that divergent selection for response to PHA-P resulted in a divergent effect on T(H)1 cell activity, resulting in altered macrophage function in chickens. Selection, based on response to PHA-P, could lead to more resistant birds or birds with an enhanced immune response.     

Identifying critical parameters in the dynamics and control of microparasite infection using a stochastic epidemiological model

A stochastic epidemic model is presented to study infection transmission dynamics, and hence epidemic severity and disease incidence, in a closed population. The aim was to understand the relative importance of various parameters that influence the dynamics of potential epidemics, particularly when the genetic mechanisms of resistance or tolerance to infection are considered. Simulations explored the effect of varying the transmission coefficient, latent period, recovery period, mortality rate, and the period of loss of immunity on overall epidemic outcomes. The critical parameters influencing the transmission of infection, and hence disease incidence, were the transmission coefficient, the latent period, and the recovery period; the period of loss of immunity had only trivial effects. Ideally, control strategies should decrease the transmission coefficient and/or increase the latent period and/or decrease the recovery period. By equating measured traits with disease transmission parameters, the model described in this paper can be used to identify which disease resistance genes or QTL will be truly effective in helping to develop disease-resistant livestock that suffer fewer epidemics and side-effects of infection. In particular, emphases should be placed on finding genes that decrease the transmission of infection, increase the latent period, or decrease the recovery period.     

Assessment of the dynamics of microparasite infections in genetically homogeneous and heterogeneous populations using a stochastic epidemic model

The aim of this paper was to explore the effect of genetic heterogeneity in host resistance to infection on the population-level risks and outcomes of epidemics. This was done using a stochastic epidemiological model in which the model parameters were assumed to be genetically controlled traits of the host. A finite locus model was explored, with a gene controlling the transmission coefficient (i.e., host susceptibility to infection) and a gene controlling the recovery period. Both genes were simulated to have 2 alleles with underlying additive or dominance inheritance and an independent assortment of alleles. The model was parameterized for a viral pig disease (transmissible gastroenteritis), and complete homogeneous mixing among genotypes was assumed. Mean population genotype dramatically affected epidemic outcomes, and subtle effects of heterogeneity on epidemic properties were also observed. Genetic variation in the transmission coefficient led to probabilities of epidemics occurring that were slightly greater than expected, but genetic variation in the recovery rate had no such effect. Epidemics were generally less severe in genetically heterogeneous populations than expected from the constituent subpopulations. Furthermore, the genotype of the initial infected animal had a marked effect on epidemic probabilities, particularly when genetic variation was for recovery rate. The results of this model provide useful information to determine the optimum population structures and to exploit genetic variation in resistance to infection. Applications of the proposed model in genetically heterogeneous populations for identifying practical disease management strategies are also discussed.     

Toxicity of a synthetic phenolic antioxidant,butyl hydroxytoluene (BHT), in vertebrate model zebrafish embryo (Danio rerio)

The synthetic antioxidant 3,5‐di‐tert‐butyl‐4‐hydroxytoluene (BHT) is widely used as an additive in the food, cosmetic and plastic industries to increase the tenability of food and plastic for the past 70 years. BHT is degraded to 3,5‐di‐test‐butyl‐4‐hydroxybenzaldehyde (BHT‐CHO) in mammals, as well as in the natural environment such as in river and water. The average daily intake of BHT for human being is estimated to be 0.3 mg/kg body weight. Even though it is considered safe for human at authorized level, but its ubiquitous presence in the aquatic environment and the controversial toxicological data are of great concern for human as well as aquatic life. The experimental findings of zebrafish embryo toxicity test (ZFET) showed that the acute toxicity of 96‐hr (LC₅₀) exposure during the embryogenic stage was found to be 4.388 mg/L and the effective concentration (EC₅₀) was 1.375 mg/L. The reduce heart rate from the sublethal concentrations indicates the chemical to be cardiotoxic but a further review is to be needed. The Teratogenic Index (TI) calculated to be 3.19, which implies the compound may be a potential teratogen in aquatic life. The findings obtained in this study will stretch more evidence regarding developmental toxicity of BHT, which will be of much importance in further risk assessment of ecotoxicological studies.     

Prevalence of Multi-Drug-Resistant Salmonella in Equids Maintained by Low Income Individuals and on Designated Equine Farms in India

We examined 872 equids (445 maintained by low-income individuals and 427 maintained on nine designated equine farms) and, using previously described methods for bacteria, isolated Salmonella from fecal samples of 59 (6.77%) animals. Of the 646 horses, 183 donkeys, and 43 mules that had feces cultured for Salmonella, 42 (6.5%), 7 (3.8%), and 10 (23.3%), respectively, were excreting Salmonella strains in feces. Six horse mares were excreting Salmonella enterica of two different serovars simultaneously. A total of 65 Salmonella enterica isolates belonged to 13 serovars, namely S. paratyphi B var Java (14), S. I. 4, 5, 12, 27: r, i: 1, 5 (11), S. Drogana (8), S. Newport (7), S. Saintpaul (5), S. Lagos (4), S. Typhimurium (5), S. Kottbus (3), S. Bovismorbificans (3), S. Dumfries (2), S. Tshiongwe (1) S. Weltevreden (monophasic) (1), and S. enterica ssp salamae (1). With Salmonella-specific polymerase chain reaction (PCR) using hisJ gene primers, 107 (12.3) fecal samples yielded a specific amplicon of 496 bp. On using PCR, prevalence of Salmonella in donkeys, horses, and mules was 4.9%, 10.8%, and 65.1%, respectively. With both methods of Salmonella detection in feces, prevalence was significantly higher in female than in male donkeys and horses. Salmonella shedding in feces was significantly higher in equids maintained by low-income people than those at designated equine farms. Almost all Salmonella isolates (63 of 65) had multiple-drug-resistance (MDR, resistance to three or more drugs). Salmonella isolates were commonly resistant to sulfamethoxazole (90.8%), tetracycline (70.8%), doxycycline (67.7%), furazolidone (66.2%), and colistin (55.4%). A few isolates had resistance to trimethoprim (3.1%), ciprofloxacin (3.1%), ceftriaxone (3.1%), ceftazidime (3.1%), cefoperazone (3.1%), chloramphenicol (4.6%), cefotaxime (6.2%), gentamicin (9.2%), ampicillin + cloxacillin (9.2%), cotrimoxazole (13.8%), kanamycin (13.8%), amoxicillin + clavulanic acid (16.9%), imipenem (16.9%), ampicillin (18.5%), amikacin (23.1%), neomycin (27.7%), nalidixic acid (33.8%), and streptomycin (36.9%). With the exception of 13 Salmonella isolates of S. Drogana (4), S. Newport (4), S. I. 4, 5, 12, 27: r, i: 1, 5 (4) and S. Kottbus (1) serovars, all had one or more than one plasmid. Molecular weight of plasmids ranged between 3 kDa and >87 kDa. One heavy plasmid (≥87 kda) was present in all the 52 plasmid-positive strains. Presence of plasmid could not be correlated with MDR in Salmonella isolates from equids.     

Replication of swine and equine influenza viruses in canine kidney cells.

Both swine and equine influenza viruses were replicated in Madin-Darby canine kidney cells. Hemagglutinin and complement-fixing antigens were synthesized at 4 to 8 hours postinfection and reached maximal synthesis at 16 to 20 hours. Neuraminidase activity increased 9- to 11-fold; 50% of the total hemagglutinin antigen was made before the infective virus progeny was detected.     

Forest-based betel leaf and betel nut People in Bangladesh： approach Lawachara National Park （LNP） farming of the Khasia indigenous to biodiversity conservation in

An exploratory survey was conducted among the Khasia living in Lawachara National Parkt （LNP） to investigate their depend- ency on the protected area （PA） for livelihoods, betel leaf hill farming, traditional forest conservation and perceptions of the PA, Field data were collected by interviewing 48 household heads from two villages （punjis） located inside the LNP and visiting their farms.： The economy of the Khasia was forest-based. They were largely dependent on betel leaf hill farming in LNP and nearly 71% of their mean annual income was de- rived from this irrespective of farmer category. On average, about 14% of the incomes of the poorer farmers came from forest produce followed by 10% and 6% for medium and rich farmers respectively. Hills and forests were the foundations of their lives and livelihoods, and LNP was the life-blood of Khasia survival. As a sustainable production system, this farming practice plays a vital role in conserving biodiversity in LNP and might be replicated elsewhere.     

Molecular characterization and xenogenic application of wharton’s jelly derived caprine mesenchymal stem cells

Aim of the present study was in vitro expansion and characterization of caprine wharton’s jelly derived mesenchymal stem cells (cWJ-MSCs) to investigate their tissue healing potential in xenogenic animal model. Plastic adherent fibroblastoid cell populations with distinctive homogeneous morphology were isolated from caprine wharton’s jelly explants. These wharton’s jelly derived cells were found positive for the surface markers CD-73, STRO-1 and CD-105, whereas they were negative for hematopoetic stem cell marker CD-34. In vitro cultured cWJ-MSCs also showed differentiation properties into osteogenic, adipogenic and chondrogenic lineages as demonstrated by von Kossa, Oil Red- O and Alcian blue staining respectively, which was further confirmed and quantified by flow cytometric analysis. Furthermore, these well characterized cWJ-MSCs were evaluated for the wound-healing potential in full-thickness skin wounds in rabbit model for 28 days. Caprine WJ- MSCs treated skin wounds showed significantly (P?<?0.05) higher percentage of wound contraction especially at the 21^st day post transplantation when compared to PBS treated control group animals. Further, we observed better healing potential of cWJ-MSCs in terms of histo-morphological evaluation, epithelialisation and collagenization with matured vascularization stage by day 28 as compared to control. In conclusion, cWJ- MSCs provide an alternative inexhaustible source of mesenchymal stem cells and also unravel new perspectives pertaining to the therapeutic use of these cells in different species.