Excessive growth of granulation tissue (EGT) is a common complication during wound healing in horses. Wounds involving lower limbs are especially prone to the development of EGT. Often increased expression of transforming growth factor-beta (TGF-beta) is found in the EGT. In the present report, resveratrol was used as TGF-beta modifier. Topical application of an aqueous solution of resveratrol at 0.01 mg/mL once daily for 5-7 weeks depending on EGT size, successfully suppressed the EGT growth in all seven horses in the present case report.

Background: Staphylococcus aureus is a prevalent opportunistic pathogen which is increasingly associated with various equine dermatological afflictions. The burgeoning issue of antibacterial resistance against this bacterium necessitates novel therapeutic approaches. This study, executed from May to November (2022) at the National Research Centre on Equine, Equine Production Centre, Bikaner, aimed to isolate and identify S. aureus from equine dermal lesions and to assess the in vitro efficacy of both organic (methanolic, aqueousand ethanolic) and inorganic (chloroform and petroleum ether) phytoextracts from Calotropis gigentean, Capparis decidua, Leptadenia pyrotechnica, Aerva javanica, Azadirachta indica, Aloe vera and Eucalyptus camaldulensis. Method: The study utilised agar well diffusion and broth dilution techniques to assess the antimicrobial efficacy of these extracts against S. aureus. Result: Microscopic analysis of gram-stained smears from cultures, alongside a suite of biochemical assays and polymerase chain reaction (PCR), corroborated the presence of S. aureus. The antimicrobial screening disclosed that both organic and inorganic extracts of E. camaldulensis manifested the most pronounced antibacterial activity, exhibiting zones of inhibition ranging from 15 mm to 21 mmand minimum inhibitory concentrations between 1.56 to 3.13 mg/mL. Furthermore, extracts from A. indica (chloroform, methanolicand ethanolic) and A. vera (methanolic and ethanolic) also demonstrated antibacterial effectiveness against this pathogen, with inhibition zones extending from 15 mm to 17.33 mm (MIC: 3.13 to 25 mg/mL) and 9 mm to 12 mm (MIC: 12.5 to 25 mg/mL), respectively. Moreover, the outcomes of this investigation substantiate the antibacterial capabilities of E. camaldulensis, A. indica and A. vera against dermatological pathogens, advocating their inclusion in topical antibacterial formulations as a strategic countermeasure to the escalating challenge of drug resistance.

The coronavirus disease 2019 (COVID-19) pandemic highlighted the critical need for broad-spectrum antivirals with high resistance barriers. Here, we demonstrate that SB431542, a selective TGF-β receptor I (ALK5) inhibitor, exhibits potent antiviral activity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) through unprecedented multitargeted mechanisms. Through comprehensive in vitro, isothermal titration calorimetry, and in silico analyses, we identified that SB431542 directly binds to SARS-CoV-2 ORF3a and disrupts its canonical function in inhibiting autophagosome-lysosome fusion. This interaction restored lysosomal acidification and normalized perinuclear LAMP-1 localization, significantly impairing virion assembly as evidenced by disrupted nucleocapsid-RNA association and reduced intracellular viral titers. Additionally, SB431542 downregulated the CLEAR network genes responsible for lysosomal biogenesis, further restricting viral egress pathways. Our temporal analyses revealed that at later infection stages (36-48 hours post-infection [hpi]), SARS-CoV-2 exploits TGF-β-induced lysosomal membrane permeabilization (LMP) and apoptosis for viral release-processes effectively inhibited by SB431542 through suppression of GADD45b and BAX expression. These multiple mechanisms resulted in an exceptional EC50 of 751.8 nM against SARS-CoV-2. In vivo efficacy was demonstrated in embryonated chicken eggs, where SB431542 conferred dose-dependent protection against lethal infectious bronchitis virus (IBV) challenge, with a favorable therapeutic index of 34.54. Remarkably, sequential passaging of SARS-CoV-2 for 50 generations under SB431542 selection pressure failed to generate resistant variants, contrasting sharply with the rapid resistance emergence typical of direct-acting antivirals. These findings establish SB431542 as a promising broad-spectrum coronavirus inhibitor with a unique triple-mechanism approach that simultaneously targets viral entry via TGF-β/Smad modulation, disrupts ORF3a-mediated lysosomal dysfunction affecting assembly, and attenuates TGF-β-induced apoptosis during late-stage infection, collectively imposing multiple selective constraints that impede escape mutation development. IMPORTANCE:The COVID-19 pandemic highlighted the urgent need for antiviral drugs with high barriers to resistance. This study reveals that SB431542, a drug previously developed to inhibit TGF-β signaling, exhibits remarkable effectiveness against SARS-CoV-2 through an unprecedented triple-mechanism approach. Unlike conventional antivirals that target a single viral component, SB431542 simultaneously disrupts viral entry, assembly, and release by binding to the viral ORF3a protein and modulating host cellular processes. Most importantly, SARS-CoV-2 failed to develop resistance against SB431542 even after 50 generations of exposure-a significant advantage over current therapeutics that quickly lose effectiveness due to viral mutations. Our findings also uncover that coronaviruses exploit both lysosomal dysfunction and programmed cell death to spread efficiently, providing new targets for therapeutic intervention. This research establishes SB431542 as a promising broad-spectrum coronavirus inhibitor and demonstrates the value of targeting host-virus interactions to overcome antiviral resistance.

Rhodococcus equi infection is considered as the most common cause of severe pneumonia in foals and has a substantial impact on equine breeding farms because of its high prevalence and case fatality rate. The objective of the study was to identify prevalence of Rhodococcus equi in foals of west north Rajasthan. Fecal swab samples were collected and cultured on Rhodococcus equi specific CAZ-NB media and DNA sample was extracted. 100 foals screened from west north Rajasthan state, out of which 18 foals were found positive based on PCR results for the Rhodococcus equi infection as these samples showed the amplification of Vap A and Vap C genes. The prevalence of Rhodococcus equi in female foals was 26 per cent while the prevalence of Rhodococcus equi in male foals was 9 per cent.

Micro RNAs (miRNAs) have been implicated in the regulation of maturation, proliferation, differentiation, and activation of immune cells. In this study, we demonstrated that miR-29a antagonizes IFN-gamma production at early times post-LSDV infection in cattle. miR-29a was predicted to target upstream IFN-gamma regulators, and its inhibition resulted in enhanced IFN-gamma production in sensitized peripheral blood mononuclear cells (PBMCs). Further, stimulation of PBMCs with LSDV antigen exhibited lower levels of miR-29a, concomitant with a potent cell-mediated immune response (CMI), characterized by an increase in LSDV-specific CD8+ T cell counts and enhanced levels of IFN-gamma, which eventually facilitated virus clearance. In addition, a few immunocompromised cattle (developed secondary LSDV infection at similar to 6 months) that failed to mount a potent cell-mediated immune response, were shown to maintain higher miR-29a levels. Furthermore, as compared to the sensitized crossbred cattle, PBMCs from sensitized Rathi (a native Indian breed) animals exhibited lower levels of miR-29a along with an increase in CD8+ T cell counts and enhanced levels of IFN-gamma. Finally, we analysed that a >= 60% decrease in miR-29a expression levels in the PBMCs of sensitized cattle correlated with a potent CMI response. In conclusion, miR-29a expression is involved in antagonizing the IFN-gamma response in LSDV-infected cattle and may serve as a novel biomarker for the acute phase of LSDV infection, as well as predicting the functionality of T cells in sensitized cattle. In addition, Rathi cattle mount a more potent CMI response against LSDV than crossbred cattle.

The experiment aimed to assess the in vitro bactericidal activity of ethanolic, chloroformic, and Sequentially Extracted Water Extract (SEWE) derived from the leaves of Bougainvillea and Ziziphus nummularia (Burm. f.) Wight & Arn. (also known as Jhar ber / jhar beri / wild jujube / Indian jujube). The ethanolic leaves extract of Bougainvillea and Ziziphus nummularia showed no action against positive Rhodococcus equi strains Vap A and Vap C in the initial screening conducted using the disc diffusion method. When compared to the antibiotics already in use, namely azithromycin and rifampicin, the ethanolic leaves extract of these plants did not exhibit any antibacterial action against R. equi in laboratory tests. Nevertheless, the ample abundance of plant leaves indicates the need for additional investigation on the antibacterial activity demonstrated in vitro using different solvents against R. equi.

Occurrence of seasonal allergic dermatitis, especially insect bite hypersensitivity and atopic dermatitis are very common in horses. At present there is no satisfactory treatment is available for management of these skin allergies. Plant flavonoids such as quercetin, kaempferol and their glycoside derivatives have been reported for their anti allergic and antiinflammatory properties. In present study we have prepared an extract from the leaves of Aerva javanica, which was found effective to manage clinical cases of seasonal allergic dermatitis in horses. Clinical signs of the allergic dermatitis in horses were alopecia, thickening of skin and itching. On histopathological examination of skin biopsy samples taken from the clinical cases epidermal hyperplasia, orthokeratotic and parakeratotic hyperkeratosis, spongiosis, occasional trichomalacia, multifocal areas of aggregation of lymphocytes with or without infiltration of eosinophils were observed. Mass spectrometer analysis of more purified extract suggested a glycoside of kaempferol may be an active ingredient of the extract.

The present study aimed to examine the protective potential of cholesterol loaded cyclodextrin (CLC) at varying concentrations against sperm morphological abnormalities of cooled and frozen thawed semen of Marwari stallions. A total of forty eight ejaculates (six stallions and eight ejaculates per stallion) were promptly processed following semen collection and subjected to gross assessment of semen quality attributes. The assessment of sperm morphologies in fresh equine semen was conducted using Eosin and Nigrosine staining technique. The semen samples from individual Marwari stallions were evenly divided into five fractions and incubated in a water bath at 37°C for 15 min after dilution with primary extender to obtain the spermatozoa concentration at 120 × 106 sperm/ml. During incubation, the fractions of every semen sample were exposed to different concentrations of CLC as 0, 1.0, 1.5, 2.0 and 3.0 mg/ml of extender (C, T1, T2,T3, and T4, respectively). All the 5 aliquots were cooled and subsequently subjected to cryopreservation. Each cooled and frozen- thawed (at 37°C for 30 sec) semen samples were analyzed to assess the morphologies of the spermatozoa. Based on the findings, it was concluded that none of the CLC treatment could render any significant protection against the sperm morphological abnormalities at either pre-freeze and/or post-thaw stage of semen samples from Marwari stallions.

In this study, we demonstrated the antiviral efficacy of hesperetin against multiple poxviruses, including buffalopox virus (BPXV), vaccinia virus (VACV), and lumpy skin disease virus (LSDV). The time-of-addition and virus step-specific assays indicated that hesperetin reduces the levels of viral DNA, mRNA, and proteins in the target cells. Further, by immunoprecipitation (IP) of the viral RNA from BPXV-infected Vero cells and a cell-free RNA-IP assay, we demonstrated that hesperetin-induced reduction in BPXV protein synthesis is also consistent with diminished interaction between eukaryotic translation initiation factor eIF4E and the 5' cap of viral mRNA. Molecular docking and MD simulation studies were also consistent with the binding of hesperetin to the cap-binding pocket of eIF4E, adopting a conformation similar to m7GTP binding. Furthermore, in a BPXV egg infection model, hesperetin was shown to suppress the development of pock lesions on the chorioallantoic membrane and associated mortality in the chicken embryos. Most importantly, long-term culture of BPXV in the presence of hesperetin did not induce the generation of drug-resistant viral mutants. In conclusion, we, for the first time, demonstrated the antiviral activity of hesperetin against multiple poxviruses, besides providing some insights into its potential mechanisms of action.

Rhodococcus equi infection is considered as the most common cause of severe pneumonia in foals and has a substantial impact on equine breeding farms because of its high prevalence and case fatality rate. The present study was conducted to identify the risk factor associated with Rhodococcus equi infection in foals. Fecal swab samples were grown on Rhodococcus equi specific CAZ-NB media. Based on PCR results 100 foals screened from north western of Rajasthan, 18 foals were found positive for the Rhodococcus equi infection as these samples showed the amplification of Vap A and Vap C genes. It was found that foals below 6 months of age, born in July and August months reared at the equine farm, having large horse population, reared on sandy soil were at higher risk for getting R. equi infection.

Colic is most important clinical problem in horses. Role of biochemical and electrolyte analysis in the diagnosis and prognosis clinical problems is very important. Present study was carried out to see the biochemical and electrolyte changes with different types of colic in indigenous horses belonging to Marwari, Zanskari and Manipuri breed of horses kept at an organised farm at Bikaner. Data of clinical cases were recorded for 8 months. Serum biochemical analyses were carried out using commercially available kits. On biochemical and electrolyte analysis no significant change was observed except few parameters. All the cases of spasmodic cases responded well to the primary treatment for colic.

In the present study, the research work was carried out during June 2018 to March 2019 at ICAR-NRCE-EPC (Indian Council of Agricultural Research, National Research Centre on Equines, Equine Production Campus), Jorbeer, Bikaner (Rajasthan), India. The study was conducted to investigate in vitro antibacterial activity of ethanolic, chloroformic and Sequentially Extracted Water Extract (SEWE) leaves extracts of Aegle marmelos, Morus alba, Nerium indicum, Cascabela thevetia (L.) Lippold, Plumeria alba L. and Azadirachta indica against Vap A and Vap C positive Rhodococcus equi. Fresh leaves of these plants were collected manually from campus of ICAR-NRCE-EPC, Jorbeer, Bikaner (Rajasthan). In initial screening ethanolic leaves extract of these plants except Azadirachta indica, were found non-active against Rhodococcus equi. Chloroformic leaves extracts of Azadirachta indica did not showed in vitro antibacterial activity against Rhodococcus equi. While ethanolic and Sequentially Extracted Water Extract (SEWE) leaves extracts of Azadirachta indica showed good in vitro antibacterial activity against Rhodococcus equi. Further, solvent based fractionation, Ethanol Soluble Fraction (ESF), Methanol Soluble Fraction (MSF) and Water Soluble Fraction (WSF) of polar compounds of SEWE did not showed in vitro antibacterial activity against Rhodococcus equi. On comparison with currently used antibiotics (azithromycin and rifampicin), required concentration of the leaves extract of Azadirachta indica was too high for their possibilities of in vivo use, so abundant availability of Azadirachta indica leaves and their activity against Rhodococcus equi suggests their potential for use as disinfectant against Rhodococcus equi.

Lumpy skin disease (LSD) was reported for the first time in India in 2019 and since then, it has become endemic. Since a homologous (LSD-virus based) vaccine was not available in the country, goatpox virus (GPV)-based heterologous vaccine was authorized for mass immunization to induce protection against LSD in cattle. This study describes the evaluation of safety, immunogenicity and efficacy of a new live-attenuated LSD vaccine developed by using an Indian field strain, isolated in 2019 from cattle. The virus was attenuated by continuous passage (P = 50) in Vero cells. The vaccine (50(th) LSDV passage in Vero cells, named as Lumpi-ProVac( Ind )) did not induce any local or systemic reaction upon its experimental inoculation in calves (n = 10). At day 30 post-vaccination (pv), the vaccinated animals were shown to develop antibody- and cell-mediated immune responses and exhibited complete protection upon virulent LSDV challenge. A minimum Neethling response (0.018% animals; 5 out of 26,940 animals) of the vaccine was observed in the field trials conducted in 26,940 animals. There was no significant reduction in the milk yield in lactating animals (n = 10108), besides there was no abortion or any other reproductive disorder in the pregnant animals (n = 2889). Sero-conversion was observed in 85.18% animals in the field by day 30 pv.

BACKGROUNDThe recovery of spermatozoa from the cauda epididymis may be the only option to obtain genetic material from elite stallions that had undergone castration or sudden death due to colic or severe injury.OBJECTIVETo evaluate two different protocols for retrieval of stallion epididymal spermatozoa and to evaluate different cryoprotectants on the freezability of the epididymal spermatozoa.MATERIALS AND METHODSSix epididymides from three stallions were collected immediately after routine castration under general anesthesia. In the first experiment, each epididymis (of two testes) of the same stallion were processed using different methods for retrieval of the epididymal spermatozoa and were pooled and cryopreserved either using 5% glycerol or 5% dimethyl formamide (DMF) as cryoprotectant. The semen quality parameters viz., progressive motility, HOST, viability and acrosome integrity were evaluated at the fresh, pre-freeze and post-thaw stages.RESULTSRetrograde method of flushing of epididymis yielded significantly (p < 0.05) higher concentration of the stallion sperm than that of the floating method. The qualitative semen parameters i.e., viability, plasma membrane integrity and acrosome integrity were found to be significantly restored using 5% DMF as cryoprotectant in comparison to when 5% glycerol was used.CONCLUSIONRetrograde flushing method of epididymis yielded significantly higher sperm concentration to that of the floating method, and 5% DMF as cryoprotectant provided acceptable freezability of stallion epididymal spermatozoa. DOI: 10.54680/fr23310110312.

In this study, hesperetin was shown to inhibit the replication of multiple poxviruses, including buffalopox virus (BPXV), vaccinia virus, and lumpy skin disease virus (LSDV). Hesperetin mainly suppressed viral protein synthesis without affecting other steps of the viral life cycle such as attachment, entry, and budding. In a chromatin immunoprecipitation (CHIP) assay, we further demonstrated that hesperetin-induced reduction in BPXV protein synthesis is due to disruption of the binding of the 5’ cap of viral mRNA with the cellular translation initiation factor eIF4E. The molecular docking and MD simulation studies, also confirmed binding of the hesperetin with the cap-binding pocket of eIF4E, in a similar conformation as m7GTP binds. In a BPXV egg infection model, hesperetin was shown to suppress the development of pock lesions on the chorioallantoic membrane, as well as the associated mortality of the chicken embryos. Most importantly, long-term culture of BPXV in the presence of hesperetin did not induce the generation of drug-resistant viral mutants. In conclusion, we for the first time demonstrated the antiviral activity of hesperetin against poxviruses, besides providing novel mechanistic insights into the antiviral action of hesperetin.

The purpose of this research was to isolate and identify Dermatophilus congolensis from skin infections in horses in Rajasthan using conventional methods and polymerase chain reaction and to assess the efficacy of some selected plants against D. congolensis using agar well diffusion and broth microdilution techniques. The presence of D. congolensis was confirmed by the "Tram track" appearance of coccoid forms in culture-stained smears as well as colony characters and biochemical tests. The existence of D. congolensis was also confirmed by the resulting 500 bp PCR amplicon. The zone of inhibition ranged from 13 mm to 21 mm, and the minimum inhibitory concentration (MIC) ranged from 1.562 mg/ml to 3.125 mg/ml for all the extracts of Eucalyptus camaldulensis when tested against D. congolensis. Zones of inhibition of 15 mm to 21 mm (MIC-3.12 5 to 6.25 mg/ml) and 11 mm (MIC-2 5 mg/ml) were seen for Azadirachta indica (chloroform, methanolic and ethanolic extract) and Aloe vera (methanolic extract), respectively, when tested on this pathogen. To combat the issue of antibiotic resistance, the results of this study suggest that E. camaldulensis, A. indica, and A. vera can be used or incorporated into topical antibacterial medicines.