Lactic acid bacteria reduce bacterial diarrhea in rabbits via enhancing immune function and restoring intestinal microbiota homeostasis.

BACKGROUND: Numerous previous reports have demonstrated the efficacy of Lactic acid bacteria (LAB) in promoting growth and preventing disease in animals. In this study, Enterococcus faecium ZJUIDS-R1 and Ligilactobaciiius animalis ZJUIDS-R2 were isolated from the feces of healthy rabbits, and both strains showed good probiotic properties in vitro. Two strains (108CFU/ml/kg/day) were fed to weaned rabbits for 21 days, after which specific bacterial infection was induced to investigate the effects of the strains on bacterial diarrhea in the rabbits. RESULTS: Our data showed that Enterococcus faecium ZJUIDS-R1 and Ligilactobaciiius animalis ZJUIDS-R2 interventions reduced the incidence of diarrhea and systemic inflammatory response, alleviated intestinal damage and increased antibody levels in animals. In addition, Enterococcus faecium ZJUIDS-R1 restored the flora abundance of Ruminococcaceae1. Ligilactobaciiius animalis ZJUIDS-R2 up-regulated the flora abundance of Adlercreutzia and Candidatus Saccharimonas. Both down-regulated the flora abundance of Shuttleworthia and Barnesiella to restore intestinal flora balance, thereby increasing intestinal short-chain fatty acid content. CONCLUSIONS: These findings suggest that Enterococcus faecium ZJUIDS-R1 and Ligilactobaciiius animalis ZJUIDS-R2 were able to improve intestinal immunity, produce organic acids and regulate the balance of intestinal flora to enhance disease resistance and alleviate diarrhea-related diseases in weanling rabbits.

Summer mortality syndrome bacterial pathogens in farmed Nile tilapia (Oreochromis niloticus).

Background: The high summer mortality in many fish farms, which had detrimental economic and social implications, was a serious challenge that the fish industry had to deal with. Aim: With an examination of the most effective antibiotic, the ongoing research was intended to shed light on the identification of the main bacterial pathogens associated with the summer mortality syndrome in the diseased farmed Nile tilapia. Methods: Six hundred dead Nile tilapia samples that had suffered from summer mortality were collected from several fish farms between May and October of 2022. The gathered fish displayed hemorrhagic areas on the skin, scale detachment, fin degeneration, erosions, skin ulcers, and corneal opacity with unilateral and/or bilateral exophthalmia. The most prominent internal appearance was swelling of the internal organs with sanguineous ascetic fluid. Results: There were 225 bacterial isolates found. Six species were identified through phenotypic and biochemical analysis; they were Aeromonas, Vibrio, Streptococcus, Pseudomonas, Enterococcus, and Edwardsiella spp., in descending percentage, respectively. Aeromonas spp., Vibrio spp., and Streptococcus spp. were the three most frequent isolated bacterial pathogens. The identification of Aeromonas hydrophila, Vibrio spp., and Streptococcus iniae, the three most common bacterial isolates, was confirmed by molecular analysis by polymerase chain reaction. Most of the tested strains were found to be responsive to Ciprofloxacin (CIP), Gentamicin (CN), and Chloramphenicol (C) but resistant to Amoxicillin (AMX), according to an antibiotic sensitivity test. Conclusion: The three most dangerous common bacterial infections discovered during mass-farmed tilapia summer mortality are A. hydrophil a, Vibrio sp., and S. iniae. This makes it clear that high water temperatures may raise the possibility of bacterial infections, which could cause widespread tilapia mortality and substantial financial losses. Therefore, it is crucial to maintain a beneficial fish culture, environment, and husbandry practices to enhance the tilapia-rearing environment and lessen the virulence of the disease. Isolated bacterial strains showed low levels of resistance to AMX but were vulnerable to CIP, CN, and C.

Laboratory tools for the direct detection of bacterial respiratory infections and antimicrobial resistance: a scoping review.

Rapid laboratory tests are urgently required to inform antimicrobial use in food animals. Our objective was to synthesize knowledge on the direct application of long-read metagenomic sequencing to respiratory samples to detect bacterial pathogens and antimicrobial resistance genes (ARGs) compared to PCR, loop-mediated isothermal amplification, and recombinase polymerase amplification. Our scoping review protocol followed the Joanna Briggs Institute and PRISMA Scoping Review reporting guidelines. Included studies reported on the direct application of these methods to respiratory samples from animals or humans to detect bacterial pathogens ±ARGs and included turnaround time (TAT) and analytical sensitivity. We excluded studies not reporting these or that were focused exclusively on bioinformatics. We identified 5,636 unique articles from 5 databases. Two-reviewer screening excluded 3,964, 788, and 784 articles at 3 levels, leaving 100 articles (19 animal and 81 human), of which only 7 studied long-read sequencing (only 1 in animals). Thirty-two studies investigated ARGs (only one in animals). Reported TATs ranged from minutes to 2 d; steps did not always include sample collection to results, and analytical sensitivity varied by study. Our review reveals a knowledge gap in research for the direct detection of bacterial respiratory pathogens and ARGs in animals using long-read metagenomic sequencing. There is an opportunity to harness the rapid development in this space to detect multiple pathogens and ARGs on a single sequencing run. Long-read metagenomic sequencing tools show potential to address the urgent need for research into rapid tests to support antimicrobial stewardship in food animal production.

Phage cocktail superimposed disinfection: A ecological strategy for preventing pathogenic bacterial infections in dairy farms.

Bovine mastitis (BM) is mainly caused by bacterial infection that has a highly impact on dairy production, affecting both economic viability and animal well-being. A cross-sectional study was conducted in dairy farms to investigate the prevalence and antimicrobial resistance patterns of bacterial pathogens associated with BM. The analysis revealed that Staphylococcus (49%), Escherichia (16%), Pseudomonas (11%), and Klebsiella (6%) were the primary bacterial pathogens associated with mastitis. A significant proportion of Staphylococcus strains displayed multiple drug resistance. The use of disinfectants is an important conventional measure to control the pathogenic bacteria in the environment. Bacteriophages (Phages), possessing antibacterial properties, are natural green and effective disinfectants. Moreover, they mitigate the risk of generating harmful disinfection byproducts, which are commonly associated with traditional disinfection methods. Based on the primary bacterial pathogens associated with mastitis in the investigation area, a phage cocktail, named SPBC-SJ, containing seven phages capable of lysing S. aureus, E. coli, and P. aeruginosa was formulated. SPBC-SJ exhibited superior bactericidal activity and catharsis effect on pollutants (glass surface) compared to chemical disinfectants. Clinical trials confirmed that the SPBC-SJ-based superimposed disinfection group (phage combined with chemical disinfectants) not only cut down the dosage of disinfectants used, but significantly reduced total bacterial counts on the ground and in the feeding trough of dairy farms. Furthermore, SPBC-SJ significantly reduced the abundance of Staphylococcus and Pseudomonas in the environment of the dairy farm. These findings suggest that phage-based superimposed disinfection is a promising alternative method to combat mastitis pathogens in dairy farms due to its highly efficient and environmentally-friendly properties.

Molecular detection of bacterial zoonotic abortive agents from ruminants in Turkey.

Abortions in cattle and sheep are one of the major causes of economic losses worldwide. Brucella spp. are the most common infectious agent associated with these abortions. However, abortions caused by bacteria such as Listeria spp., Leptospira spp., Campylobacter spp. and Mycoplasma spp. are usually overlooked due to their sporadic nature and their status as non-priority abortion agents. In our study, we investigated the bacteria associated with abortion cases in cattle and sheep using PCR. For this purpose, we collected vaginal swab samples (n: 110) of aborted cattle and sheep, as well as stomach content samples (n: 69) of aborted calves and lambs from various cities in Turkey. The samples were analysed by bacteria-specific PCR to detect Campylobacter fetus, Leptospira spp., Listeria spp., Mycoplasma spp., and Yersinia spp. PCR analyses revealed that the investigated bacterial agents were present in 18.85% and 19.3% of the cattle and sheep samples, respectively, with an overall percentage of 18.99%. While the overall positivity rate for C. fetus, Leptospira spp., and Mycoplasma spp. was 2.79%, 10.06%, and 4.47%, respectively, the positivity rate for co-infection with Leptospira spp. and C. fetus was 1.68%. All samples were found to be negative for Yersinia spp. and Listeria spp. The high C. fetus positivity rate detected in sheep and in the stomach contents was statistically significant (p < 0.05). However, the difference in positivity rates between the cities, hosts, co-infections and causative agents was statistically insignificant (p > 0.05). This study provides preliminary data on the significant involvement of C. fetus, Leptospira spp. and Mycoplasma spp. in cattle and sheep abortions in Turkey indicating that they should not be overlooked in diagnosis. In addition, further research is needed to investigate the zoonotic potential of these pathogens for public health in Turkey.

Unraveling frontiers in poultry health (part 1) - Mitigating economically important viral and bacterial diseases in commercial Chicken and Turkey production.

This symposium offered up-to-date perspectives on field experiences and the latest research on significant viral and bacterial diseases affecting poultry. A highlight was the discussion on the use of enteroids as advanced in vitro models for exploring disease pathogenesis. Outcomes of this symposium included identifying the urgent need to improve the prevention and control of avian influenza by focusing research on vaccine effectiveness. In this regard, efforts should focus on enhancing the relatedness of vaccine antigen to the field (challenge) virus strain and improving immunogenicity. It was also revealed that gangrenous dermatitis could be controlled through withholding or restricting the administration of ionophores during broiler life cycle, and that administration of microscopic polymer beads (gel) based-live coccidia vaccines to chicks could be used to reduce necrotic enteritis-induced mortality. It was emphasized that effective diagnosis of re-emerging Turkey diseases (such as blackhead, fowl cholera, and coccidiosis) and emerging Turkey diseases such as reoviral hepatitis, reoviral arthritis, Ornithobacterium rhinotracheale infection, and strepticemia require complementarity between investigative research approaches and production Veterinarian field approaches. Lastly, it was determined that the development of a variety of functionally-specific enteroids would expedite the delineation of enteric pathogen mechanisms and the identification of novel vaccine adjuvants.

Usefulness of combining computed tomography and air sac fluid examination to rule out aspergillosis: case study in two gentoo penguins (Pygoscelis papua) with respiratory clinical signs.

Pulmonary aspergillosis occurring in captive penguins living in zoos and aquariums is a fatal disease owing to its high mortality rate. On conducting computed tomography, two gentoo penguins (Pygoscelis papua) with respiratory clinical signs that were housed at an aquarium in Hokkaido, Japan were found to have a certain amount of fluid in their air sacs. This fluid was collected and cultured for bacteria and fungi, the results of which both came back negative. These results enabled us to rule out bacterial infection or aspergillosis and supported our decision not to administer antibacterial and antifungal drugs. Overall, the combination of computed tomography and air sac fluid examination was useful for ruling out bacterial infection or aspergillosis in penguins with respiratory clinical signs.

Severe bronchiectasis resulting from chronic bacterial bronchitis and bronchopneumonia in a jungle cat.

Bronchiectasis is irreversible bronchial dilation that can be congenital or acquired secondary to chronic airway obstruction. Feline bronchiectasis is rare and, to our knowledge, has not been reported previously in a non-domestic felid. An ~10-y-old female jungle cat (Felis chaus) was presented for evaluation of an abdominal mass and suspected pulmonary metastasis. The animal died during exploratory laparotomy and was submitted for postmortem examination. Gross examination revealed consolidation of the left caudal lung lobe and hila of the cranial lung lobes. Elsewhere in the lungs were several pale-yellow pleural foci of endogenous lipid pneumonia. On cut section, there was severe distension of bronchi with abundant white mucoid fluid. The remaining lung lobes were multifocally expanded by marginal emphysema. Histologically, ectatic bronchi, bronchioles, and fewer alveoli contained degenerate neutrophils, fibrin, and mucin (suppurative bronchopneumonia) with rare gram-negative bacteria. Aerobic culture yielded low growth of Proteus mirabilis and Escherichia coli. There was chronic bronchitis, marked by moderate bronchial gland hyperplasia, lymphoplasmacytic inflammation, and lymphoid hyperplasia. The palpated abdominal mass was a uterine endometrial polyp, which was considered an incidental, but novel, finding. Chronic bronchitis and bronchopneumonia should be considered as a cause of bronchiectasis and a differential diagnosis for respiratory disease in non-domestic felids.

A Review of the Industrial Importance, Common Bacterial Diseases, and Zoonotic Risks of Freshwater Aquarium Fish.

Background: The ever-increasing popularity of home aquariums, most often involving freshwater varieties, has exploded in recent years partially due to the Coronavirus pandemic and related to stay-at-home public health precautions for social distancing. With this ever-increasing popularity of aquariums as a hobby, and whether this involves freshwater or marine fish species, a number of important economic, ecological, and public health issues arise for both fish and hobbyists alike. Materials and Methods: This review highlights the history and genesis of aquariums as both a hobby and an important economic factor (industrial, commercial) for many countries on a global basis. Types of aquarium fish are described, and culture conditions leading to homeostasis in aquatic environments are detailed. When these conditions are not met and aquatic systems are out of balance, the disease can result due to stressed fish. Results: Major bacterial diseases associated with freshwater aquarium fish are reviewed, as are potential human infections related to the care and maintenance of home aquaria. Conclusion: Besides, scientific information was also combined with the false facts of hobbyists who tried to identify and treat diseases during an outbreak in the aquarium. Finally, unresolved issues and important misconceptions regarding the field are discussed.

Vaginal aerobic bacteria of healthy bitches and those with fertility problems.

The most common problems in veterinary practice in bitches are bacterial infections of the reproductive tract associated with fertility problems. Research to determine the correlation between the health status of female dogs and bacterial flora of the genital tract has been ongoing for years, but the results obtained by different authors are often contradictory, and do not always concern breeding bitches. Our study identified the most common aerobic bacteria in the genital tract of numerous breeding bitches population. A total of 275 breeding dogs in anestrous phase of the estrous cycle were included in this study. 198 were qualified to the first group with no genital tract infections and no reproductive disorders. 68 bitches were qualified to the second group with complications such as: infertility, abortion, foetus resorptions and newborn mortality. The type of bacterial isolates was almost the same in the healthy bitches and the group with fertility problems. The most common bacteria obtained from the vaginal tract of the tested dogs were Streptococcus spp., Staphylococcus spp., Mycoplasma canis and Escherichia coli. There were no significant differences in bacterial prevalence in the group with reproductive problems versus healthy dogs; however, we found a statistically significant difference between both groups when the numbers of bacterial strains were compared. The number of one-strain bitches was statistically higher in the problematic group than in the non-problematic one. Bacterial culturing of vaginal swab specimens from breeding bitches without clinical signs of genital disease is of little value. Furthermore, it should always be preceded by an examination (clinical, cytological or vaginoscopy etc.). The request or requirement to perform vaginal cultures that is made by some breeders, while common, is not diagnostic for any pathologic condition and the results of these cultures should never be used to determine if antibiotic therapy is indicated.

Treatment of postpartum endometritis induced by multidrug-resistant bacterial infection in dairy cattle by green synthesized zinc oxide nanoparticles and in vivo evaluation of its broad spectrum antimicrobial activity in cow uteri.

Postpartum endometritis significantly affects the health and productivity of cattle, causing significant economic loss that is speculated to exceed billions of dollars annually. Treatment of postpartum endometritis, which is linked to various bacterial infections in the uterus after delivery and has an alarmingly high risk of antibiotic treatment failure for unidentified reasons, represents a great challenge. Several studies have demonstrated that various disease complications, such as multidrug-resistant (MDR) bacterial strains, prolonged infection treatment, and increased mortality risk, have emerged as a result of the extensive use of antibiotics to treat uterine infections and other microbial-related diseases. Recent research has led to the development of zinc oxide nanoparticles (ZnO NPs) that exhibit broad-spectrum antibacterial efficacy against bacterial pathogens, including MDR bacteria, without producing mutants that are resistant to zinc oxide (ZnO). In the present work, we biologically synthesized ZnO NPs from a green natural source of Helianthus annuus seeds for the treatment of endometritis caused by MDR bacterial strains in dairy cattle. We examined ZnO's potential as a substitute antimicrobial agent to treat cow endometritis by testing its ability to sustain potent antimicrobial activity against pathogenic bacteria, including Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus), in cow uteri. Among uterine bacteria, ZnO significantly decreased E. coli and S. aureus, which are known pathogenic bacteria within the uterus and achieved a high cure rate that was associated with the induction of estrous and pregnancy. Taken together, our observations of ZnO's broad range of antibacterial activity in-vivo with an animal model and subsequent evaluations of its therapeutic efficacy in cows with endometritis shed light on its potential to be used as a substitute antimicrobial agent for the treatment of uterine illness.

Neglected bacterial infections associated to bovine respiratory disease in lactating cows from high-yielding dairy cattle herds.

Bovine respiratory disease (BRD) is a multifactorial and predominantly multietiological disease that affects dairy cattle herds worldwide, being more frequent in young animals. The occurrence of BRD was investigated in lactating cows from two high-yielding dairy herds in southern Brazil. To determine the etiology of the clinical cases of acute respiratory disease, nasal swab samples were collected from cows with clinical signs of BRD and evaluated using PCR and RT-PCR for nucleic acid detection of the main BRD etiological agents, including Mycoplasma bovis, Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, bovine respiratory syncytial virus, bovine coronavirus, bovine viral diarrhea virus, bovine alphaherpesvirus 1, and bovine parainfluenza virus 3. Only three microorganisms (M. bovis, H. somni, and P. multocida) were identified in both single and mixed infections. We concluded that 40.0% of the cows were infected with M. bovis and 75.0% with H. somni in herd A. Considering both single and mixed infections, the analyses performed in herd B showed that 87.5%, 25.0%, and 50.0% of the cows were infected with M. bovis, H. somni, and P. multocida, respectively. M. bovis and H. somni are considered fastidious bacteria and laboratory diagnosis is neglected. Subsequently, most clinical cases of mycoplasmosis and histophilosis in cattle remain undiagnosed. This study demonstrates the importance of M. bovis and H. somni infections in adult cows with BRD. These results highlight the importance of including these bacteria in the group of etiological agents responsible for the occurrence of BRD in cattle, especially in adult cows with unfavorable immunological conditions, such as recent calving and peak lactation.

Elution profiles of metronidazole from calcium sulfate beads.

BACKGROUND: Antibiotic beads are used to treat local bacterial infections by delivering high drug concentrations to infected tissue. OBJECTIVES: This study examined the elution characteristics of metronidazole from metronidazole-calcium sulfate (MCa) and metronidazole-calcium-potassium sulfate (MCaK) beads over 20 days and the antibacterial efficacy of the beads after storage. METHODS: The MCa and MCaK beads were prepared by mixing 250 mg of metronidazole and 10 g of calcium sulfate hemihydrate with water and a 3% potassium sulfate solution, respectively. The beads were placed in phosphate-buffered saline for the elution study. The metronidazole eluents were determined using high-performance liquid chromatography. The microstructures were examined by scanning electron microscopy (SEM), and the antimicrobial activity was evaluated by a microbioassay. RESULTS: For the 20-day study, the total amount of metronidazole released was greater in the MCa beads than in the MCaK beads by 6.61 ± 0.48 mg (89.11% ± 3.04%) and 4.65 ± 0.36 mg (73.11% ± 4.38%), respectively. The amounts of eluted drugs from the MCa and MCaK beads were higher than the minimum inhibitory concentration at 0.5 µg/mL against anaerobic bacteria at both 20 days and 14 days. SEM showed that calcium crystals on the outer surface had dissolved after elution, and thinner calcium crystals were prominent in the MCaK beads. The MCa and MCaK beads exhibited antibacterial activity after setting, followed by storage at room temperature or 4°C for 21 days. CONCLUSIONS: The MCa beads could release more drug than the MCaK beads, but all eluted metronidazole amounts were effective in controlling bacterial infections. Both metronidazole beads could be stored at ambient temperature or in a refrigerator.

[Recurrent vaginitis and cystitis due to a vagina and cervix duplex in a bitch - a case report]. / Rezidivierende Vaginitis und Zystitis aufgrund einer Vagina und Zervix duplex bei einer Hündin ­ ein Fallbericht.

Incomplete merging of the Müllerian ducts may result in vaginal septa or even completely separate vaginal canals. It may also cause cervical malformation. In most cases, there are only rudimentary vaginal septations. Only a few reports describing completely separated vaginal canals (vagina duplex) and a cervix duplex in dogs have been published. This article reports the case of a bitch presented with recurrent vaginitis and cystitis. The symptoms always appeared following the dog's heat. The bitch had been pre-treated by a prior veterinarian as well as in a prior clinic. When presented in the clinic, a vaginal septum was suspected based on digital vaginal examination. Ultrasonographic examination and urinalysis allowed for the diagnosis of bacterial cystitis, which was subsequently treated with antibiotics. The bitch was presented again for vaginal endoscopy, transection of the vaginal septum, and ovariohysterectomy. Both vagina duplex and cervix duplex were diagnosed. Due to the extensive dimensions of the findings, the altered tissue was not resected. No more bacteria were detected in a repeated urine analysis. After spaying, the bitch showed no recurrence of symptoms of cystitis or vaginitis.

Serum/Faecal S100A12, CRP and lactoferrin can be used to distinguish infectious and non-infectious canine diarrhoea.

OBJECTIVES: This study aimed to evaluate the diagnostic efficacies of selected inflammatory and intestinal biomarkers in cases of infectious and non-infectious diarrhoea in dogs. METHODS: A total of 60 dogs, 12 healthy (Control Group) and 48 with diarrhoea were used. Viral, Bacterial, Parasitic (infectious) and Nutritional diarrhoea (non-infectious) subgroups (n: 12) were formed according to the aetiology, on the basis of clinical and laboratory examinations. Selected inflammatory and intestinal biomarkers (Calgranulin, S100A12; Lactoferrin, LCTF; C-reactive protein, CRP) were measured both in serum and faecal samples. RESULTS: Compared to the Control and Nutritional Diarrhoea groups, the infectious diarrhoea groups had higher serum S100A12, LCTF, CRP, blood urea nitrogen, creatinine (CR), alanine transaminase and alkaline phosphatase, and lower glucose (GLU), sodium (Na) and potassium (K) concentrations (p < 0.05); Viral and Parasitic Diarrhoea groups had lower serum albumin (ALB) and total protein (TP) concentrations (p < 0.05). Faecal S100A12, LCTF and CRP concentrations were higher in infectious diarrhoea groups compared to the Control and Nutritional Diarrhoea groups (p < 0.05). Faecal LCTF and CRP concentrations were higher in the Bacterial Diarrhoea group than in the Viral and Parasitic Diarrhoea groups (p < 0.05). CLINICAL SIGNIFICANCE: It was determined that serum (area under curve, AUC: 0.842 and 0.956) and faecal (AUC: 0.975 and 0.786) S100A12 and CRP concentrations in viral diarrhoea; serum (AUC: 0.956) and faecal (AUC: 0.992) LCTF concentrations in bacterial diarrhoea have diagnostic values in the diagnosis of the presence of intestinal inflammation and damage and can be used in the differential diagnosis of infectious and non-infectious diarrhoea.

Identification and functional analysis of circRIPK2 in lipopolysaccharide induced chicken macrophages.

1. It was hypothesised that a circular RIPK2 (circRIPK2) highly expressed in chicken macrophages plays an important role during bacterial infection.2. After PCR amplification, Sanger sequencing and RNase R exonuclease treatment of chicken macrophages, it was found that circRIPK2 was a stable circular RNA, which was formed by reverse splicing of exons 4 to 9 of the RIPK2.3. The circRIPK2 can promote the lipopolysaccharide (LPS) induced cellular injury by reducing cell viability and increasing the expression of pro-inflammatory cytokines and apoptosis genes.4. Six miRNAs were identified as interacting with circRIPK2, potentially targeting 1,817 genes, which were significantly enriched in the Wnt signalling pathway, adherens junction and NOD-like receptor signalling pathway.5. This study provides better understanding of the function of circRIPK2, which may prove a potential biomarker and indicate potential targets for the treatment of bacterial infection.

Epizootological monitoring of some bacterial infectious diseases of animals on the territory of the Republic of Kazakhstan.

The presence of significant areas used for pastures in Kazakhstan makes it relevant to study the epizootic situation in terms of the incidence of animals during their grazing. The most frequent and dangerous diseases during this period are clostridiosis and listeriosis, which cause significant damage to livestock. Monitoring studies of the dynamics of morbidity among animals for infectious diseases during the grazing period were carried out on the basis of veterinary reporting. Bacteriological studies were carried out in a state laboratory using generally accepted microbiological methods. Analytical and statistical methods were used during the work. Monitoring studies indicate a significant spread of clostridiosis in the territory of Kazakhstan. The highest incidence rate was characterised by emphysematous carbuncle infection, which was recorded throughout the country, with an average frequency of about 40 cases per year. The incidence of braxy and anaerobic enterotoxemia occurred much less frequently and mainly in the southern regions of the country. Listeriosis was also observed only in some regions, but at the same time, a stationary focus of the disease was recorded in the Utylaut region, where cases of the disease are recorded annually. Conducted research made it possible to form the dynamics of the incidence of animals in the pasture period for these diseases, to determine the regions of the country with different intensity of the epizootic situation, as well as to carry out bacteriological isolation of a pure culture of the pathogen from samples taken in the foci of the disease. The studies made it possible to collect information on the features and mechanisms of the occurrence of diseases in the conditions of different regions of Kazakhstan and to develop a multi-vector strategy for preventive measures to eliminate animal diseases during the grazing period.

Alternatives to animal models to study bacterial infections.

Animal testing has made a significant and unequalled contribution to important discoveries and advancements in the fields of research, medicine, vaccine development, and drug discovery. Each year, millions of animals are sacrificed for various experiments, and this is an ongoing process. However, the debate on the ethical and sensible usage of animals in in vivo experimentation is equally important. The need to explore and adopt newer alternatives to animals so as to comply with the goal of reduce, refine, and replace needs attention. Besides the ever-increasing debate on ethical issues, animal research has additional drawbacks (need of trained labour, requirement of breeding area, lengthy protocols, high expenses, transport barriers, difficulty to extrapolate data from animals to humans, etc.). With this scenario, the present review has been framed to give a comprehensive insight into the possible alternative options worth exploring in this direction especially targeting replacements for animal models of bacterial infections. There have been some excellent reviews discussing on the alternate methods for replacing and reducing animals in drug research. However, reviews that discuss the replacements in the field of medical bacteriology with emphasis on animal bacterial infection models are purely limited. The present review discusses on the use of (a) non-mammalian models and (b) alternative systems such as microfluidic chip-based models and microdosing aiming to give a detailed insight into the prospects of these alternative platforms to reduce the number of animals being used in infection studies. This would enlighten the scientific community working in this direction to be well acquainted with the available new approaches and alternatives so that the 3R strategy can be successfully implemented in the field of antibacterial drug research and testing.

Identification and analysis of pathogenic bacteria causing outbreaks in Indian major carp aquaculture of Tripura.

OBJECTIVE: The objective of this study was to investigate bacterial disease outbreaks in Indian major carp from aquaculture systems in Tripura, India, and identify the bacterial species associated with those outbreaks. METHODS: A 3-year surveillance was conducted in eight districts of Tripura, during which nine bacterial disease outbreaks were recorded. Fourteen bacterial strains isolated from diseased Indian major carp were selected and identified using phenotypic, molecular (16S ribosomal RNA gene), and phylogenetic analyses. In vitro pathogenicity studies were performed to assess the potential pathogenicity of the isolated bacteria. RESULT: The selected isolated strains were preliminarily identified under the genera Aeromonas (9 isolates), Acinetobacter (1 isolate), Citrobacter (3 isolates), and Pseudomonas (1 isolate). Molecular and phylogenetic analyses confirmed the species of the isolated bacteria, including Aeromonas jandaei (strains COF_AHE09 and COF_AHE61), Aeromonas veronii (strains COF_AHE13, COF_AHE52, COF_AHE55, COF_AHE56, and COF_AHE62), Aeromonas hydrophila (strains COF_AHE51 and COF_AHE58), Acinetobacter pittii (strain COF_AHE14), Citrobacter freundii (strains COF_AHE20, COF_AHE57, and COF_AHE59), and Pseudomonas aeruginosa (strain COF_AHE54). Behavioral and clinical signs observed in the diseased fish, such as lethargy, skin hemorrhaging, ulcers, fin and tail rot, exophthalmia, distended abdomen, scale loss, and skin discoloration, indicated the presence of bacterial septicemia. The in vitro pathogenicity studies highlighted the potential role of these bacteria in disease development, especially under environmental stress. CONCLUSION: This study provides valuable insights into the diversity of bacterial species associated with bacterial disease outbreaks in Indian major carp from aquaculture systems in Tripura. It serves as the first comprehensive investigation of its kind, contributing to our understanding of bacterial infections in Indian major carp.