Background: The dairy sector faces formidable obstacles in the early detection of the bacteria causing subclinical mastitis and in the proactive treatment of those cases. The objective of the study was to access multiplex PCR assays (MPCR) and conventional methods for the concurrent identification of significant bacteria that cause sub-clinical mastitis and to compare these methods. 200 samples of pooled milk from cows and buffaloes in Rajasthan were gathered between 2020 and 2021.&nbsp; 
 
Methods: The primary bacterial pathogens in milk samples were identified using conventional methods such multiplex PCR, biochemical testing and culture.&nbsp; 
 
Result: From 200 combined samples of cow and buffalo milk, the traditional approach was able to extract 97 different strains. Staphylococcus aureus 54 (27%), Streptococcus spp. 30 (15%) and E. coli 13 (6.5%) were shown to be prevalent as single or mixed infections, respectively. Staphylococcus was the main pathogen that was discovered. concurrently, S. aureus, Streptococcus and E. coli. Direct detection of Staphylococcus 65 (32.5%), Streptococcus 37 (18.5%), E. coli and 16 (8%) by multiplex PCR was found in milk samples. The analysis revealed that because multiplex PCR assays have higher specificity and sensitivity than conventional procedures, they are more reliable. The multiplex PCR method employed in the current study was a simple and quick technique to identify the major pathogens and it has the potential to be a very helpful tool for determining the pathogens that cause environmental mastitis and evaluating the health of the herd.

Molecular Detection of Some Most Important Pathogenic Bacteria by Multiplex PCR for Diagnosis of Subclinical Mastitis in Bovine

A study was carried out to assess the efficacy of purified antimicrobial peptides of lactic acid bacteria (LAB), isolated from partially decayed samples of food items, against selected clinical isolates of mastitis bacterial pathogens, viz., Escherichia coli (E.coli), Staphylococcus aureus (S.aureus) and Streptococcus agalactiae (S.agalactiae). The crude culture extracts (CCE) from LAB isolates were concentrated and purified by salting out method followed by dialysis. Partially purified peptides (Tp-5 CRp-2 CRp-3) from three LAB isolated from tomato and carrot showed a significantly higher inhibitory spectrum, with zone of inhibition ranging from 11.66&plusmn;0.33 to 12.66&plusmn;0.33 mm diameter against different test pathogens. The inhibitory efficiency of all the three peptides was found to be significantly superior to the peptides from reference strain Lactococcus lactis NCIM-2114. Minimum inhibitory concentration (MIC) of the purified peptide samples ranged between 0.79&plusmn;0.09 to 1.11&plusmn;0.09 &micro;g/ml against different test pathogens. The activity of peptides was completely lost after treatment with protease, thereby confining their protein character.&nbsp;

Efficacy of purified antimicrobial peptides from lactic acid bacteria&nbsp;against bovine mastitis pathogens

<div>Background: Despite the improved modern technologies to inactivate pathogens, there has been a continuous surge of food-borne diseases involving many bacterial pathogens like Staphylococcus aureus, Escherichia coli, Bacillus cereus, Salmonella, Campylobacter, Listeria etc. Physical methods of reduction of microbial load in raw foods have been known to negatively impact the organoleptic properties of the products hence reducing their acceptability. Therefore, knowing that food may be laden with many pathogens, the biocontrol methods may be an effective alternative to chemical and physical methods of preservation for eliminating pathogens. 
 
Methods: The pathogens to be tested in the experiment were recovered from various food sources and clinical samples viz., milk, raw chicken and a faecal sample, chicken. For evaluation of inhibitory characteristics of Lactobacillus spp. (Lactobacillus plantarum) against selected pathogenic bacteria the grown culture of pathogens (S. aureus, E. coli, B. cereus) was added with Lactobacillus spp. at different concentrations and incubated at 24, 36 and 48 hrs and mean CFU units were evaluated. All of the treated groups were compared with the concurrent untreated groups of respective bacteria. 
 
Result: After 24 hrs of incubation at 37&deg;C, the mean colony forming units (CFU/ml) of S. aureus, E. coli, B. cereus in the cultured groups treated with 108, 107, 106, 105 and 104/ml of L. plantarum were 2.56 x 108, 2.78 x 108, 2.90 x 108, 3.03 x 108 and 39.19 x 108; 0.81 x 108, 1.09 x 108, 1.36 x 108, 1.85 x 108 and 24.62 x 108; 0.31 x 108, 0.43 x 108, 0.60 x 108, 0.69 x 108 and 7.12 x 108, respectively. Henceforth, the mean colony forming counts of all the three pathogens increased when the concentration of L. plantarum decreased.</div>

Evaluation of Efficiency and Extent of Inhibitory Characteristics of Lactobacillus spp. (Lactobacillus plantarum) against Selected Pathogenic Bacteria

The study was conducted to observe the individual and synergetic effect of certain free-living diazotrophic bacteria, viz., Azospirillum brasilense, Azotobacter chroococcum, Clostridium pasteurianum; a P-solubilising bacteria, viz., Pseudomonous denitrificans and Rhizobium on growth of rice cultivar NDR &ndash; 359. The resuts showed that all the inoculated strains both individually and in combinations played a positive role in enhancement of growth of rice cultivar. It was also observed that multi-strain inoculation responded better, which indicated their synergetic effect.

Synergetic effect of different free-living diazotrophic bacteria, P &ndash; solubilizing bacteria and Rhizobium on growth of Oryza sativa L. (cv. NDR &ndash; 359)

<div>Nematodes are the most abundant metazoans on the planet can be found in almost all the ecosystems on earth. They may be free-living, marine, bacterivorous, or parasites of plants and animals, including human beings. Nematodes always interact with other organisms, including- viruses, fungi, protozoa, rickettsia, tardigrades and bacteria in their microbiome. Nematodes are particularly well suited for investigating host associations with bacteria because they have a long history of evolution. Nematode-bacteria interaction is now being used as a model for investigating animal-microbe interaction and can be positive (mutualistic) or negative (pathogenic/parasitic) and may be transient or stably maintained (symbiotic). In any disease complex, nematodes play important roles in association with bacteria to aggravate disease intensity. Bacteria is a potential food source for nematodes and pathogenic to nematodes. Bacterial antagonists are now getting importance as potential biocontrol agents in modern agriculture due to gradual phasing out of chemical nematicides. In addition to trophic and pathogenic interactions, bacteria can serve as mutualists by aiding nematodes in development, defense, reproduction and nutrient acquisition. Furthermore, since many mechanistic aspects of nematode-bacterium interactions are conserved, their study can provide broader insights into other types of associations, including those relevant to plant and human diseases.</div>

​Current Status of Nematode-bacteria Interaction: A Mini Review

<div style="text-align: justify;">Environment is an important aspect of plant ecology and global environmental change is of major concern that is caused by natural and human activities which alter greenhouse gas concentrations in the atmosphere. The increase in concentration of greenhouse gases is foreseen to continue to raise average global temperature. Elevated concentration of carbon dioxide with increased temperature influences the plant-disease interactions. The environment has an influence over the development as well as temporal and spatial dissemination of plant diseases. The result of change in environment can either be favorable, non favorable or impartial, as these changes can either lessen, expand or have no influence on diseases as each disease may be attributed differently to these variations in accordance to an area or time of year. Variation in environmental conditions is said to be influencing plants in natural ecosystems all around the world and change in climate directly impacts crops, along with their synergy in accordance with the microbial population. The important elements governing magnification and spread of plant diseases are temperature, moisture, light and carbon dioxide concentration. Environment change causes a significant impact on germination, reproduction, sporulation, spore dispersal, along with perforation by pathogens as a vulnerable host will not be invaded by a pernicious pathogen if the environmental conditions are not facilitative for disease. The environment influences all life stages of host as well as that of a pathogen and as a result induces an opposition to various pathosystems. Resistance mechanisms of plants, including effector-triggered immunity (ETI), pattern-triggered immunity (PTI) and defence network of hormones, are particularly influenced by environmental elements. Pathogenic virulence mechanisms like fabrication of virulence proteins and toxins, and also spore germination and survival are governed by factors such as atmospheric carbon dioxide, temperature and humidity. A large number of in-vitro experiments to understand interactions between plants and pathogens rely on predetermined pathosystems by making use of ascertained environmental conditions which take into consideration relatively small part of vital plant&ndash;pathogen&ndash;environment interactions occurring in natural ecosystem. Thus, there is a need to research effectiveness of disease management strategies that is, to assess current management strategies to understand multifaceted nature of environment-pathogen interactions for production of crops that are irrepressible to environment change.</div>

Environment-Pathogen Interaction in Plant Diseases

<div>Background: Black pod rot disease of cocoa caused by (Phytophthora palmivora Butl.) is one of the major diseases on cocoa plantations worldwide. Many attempts have been made to prevent or reduce the infection of pathogens, but they have not provided optimum results. This study aims to detect antibiotic genes in endophytic bacteria that can suppress cocoa black pod disease. 
 
Methods: Eight endophytic bacteria were isolated from healthy cocoa pods and twigs that showed potentials in suppressing P. palmivora growth in vitro were used in suppressing of black pod rot disease in vivo tests. Antibiotic biosynthesis-related genes from eight endophytic bacterial isolates were confirmed by using PCR method, which includes phenazine-1-carboxylic (PCA), pyrrolnitrin (PRN), phenazine-1-carboxamide acid (PCN), pyoluteorin (PLT) and 2,4-diacetylphloroglucinol (DPAG/Phl). 
 
Result: The endophytic bacteria, 4RSI, 5BR B3 and 2RW B2 isolates showed the highest disease suppressing index to black pod rot disease in vivo, i.e., 70.27%, 70.08% and 56.64%. The isolates 5BR B3 and 2RW B2 DNA yielded PCR product by using PCA primers (1400 bp), PRN primers (700 bp) and DAPG primers (1600 bp), while the 5RSI isolate yield PCR product using PRN primers only. Endophytic bacterial isolates 5BRB3 and 2RW B2 provided partial disease suppression to pod rot disease by inhibit pathogen growth and antibiotic compounds production.</div>

​Control Activity and Antibiotic Gene Detection of Endophytic Bacteria in Suppressing Cocoa Black Pod Disease (Phytophthora palmivora Butl.)

<div style="text-align:justify">Background: Lactic acid bacteria are regarded the most important bacteria concerning food fermentation, pharmaceutical and special dietary applications. Strains have been isolated from environments rich in available carbohydrate substrates, such as food and feed, but also in human and animal cavities and in sewage and plant material. Besides lactic acid, other side products include acetate, ethanol, CO2, formate and succinate. The most important advantage of Lactic acid bacteria making them suitable for the use in food biotechnology, is that they are generally recognized as safe (GRAS). Studies on Lactic acid bacteria isolated from aerial parts of plants are scarce. Bacteria that colonize plant roots and promote plant growth are referred to as plant growth-promoting rhizobacteria. Several substances produced by antagonistic rhizobacteria have been related to pathogen control and indirect promotion of growth in many plants. The present work explores the potential use of Lactic acid bacteria in promotion of plant growth.&nbsp; 
Methods: Three isolates were obtained from aerial parts of pomegranate plant and confirmed by 16S rRNA sequencing to belong to Leuconostoc sp. The isolates were checked for plant growth promoting traits viz. antifungal activity, production of plant growth hormones, enzymes and 1-amino cyclopropane carboxylate deaminase activity.&nbsp; 
Result: As LAB showed plant growth promoting traits, they can be suitably used for plant growth promotion.</div>

​Potential of Lactic Acid Bacteria in Plant Growth Promotion

<div>Background: Mastitis, the inflammation of parenchyma of mammary gland is frequently considered to be costliest and complex disease prevalent in India. Mastitis is caused by pathogens like Staphylococcus spp., Streptococcus spp., Mycoplasma bovis, E. coli, Klebsiella spp., Citrobacter spp., Enterobacter spp. and Entercoccus. The treatment of mastitis in animals is carried out using antibiotics. Treatment failure in mastitis is due to increased antibiotic resistance of mastitis pathogens and also due to indiscriminate use of antibiotics without testing in vitro antibiotic sensitivity test against causal organisms. In comparison to cultural method, PCR assays takes less time for detection of bacteria from the mastitis milk samples. Present research work was carried out regarding isolation, identification and multiple drug resistance profile of clinical bovine mastitis associated pathogens using conventional as well as molecular approach.&nbsp; 
 
Methods: In the present study, 73 mastitis milk samples were collected from Anand and Panchmahal district of Gujarat. The milk samples were subjected for cultural isolation and DNA extraction for identification of bacteria by cultural and PCR method. Antimicrobial sensitivity pattern of the isolates were carried by disc diffusion method and isolates were categorized in multiple drug resistant. &nbsp; &nbsp;&nbsp; 
 
Result: In the present study, Out of 73 mastitis milk samples collected from cows 48 (65.75%) cows were positive for bacterial isolation and S. aureus was the most predominant bacterial species. PCR from the mastitis milk additionally detected bacteria in culturally negative milk samples. Most sensitive drug was gentamicin and most of the isolates (90.19%) showed the multiple drug resistance for the two to nine drugs with 0.1 to 0.6 multiple antibiotic resistance index.</div>

<div>Isolation, Molecular Identification and Multidrug Resistance Profiling of Bacteria Causing Clinical Mastitis in Cows</div>

<div>Background: There is scanty literature available about concurrent infection of Pestes des Petits (PPR) and emerging opportunistic bacterial pathogens in field conditions. Thus, study was conducted to investigate the above based on necropsy.&nbsp; 
 
Methods: Systematic necropsy examination was conducted on 50 small ruminant carcasses. Detailed study included virological, bacteriological, gross and microscopic investigation on tissues collected using standard techniques. PPR virus (PPRV) antigen was detected in various samples using sandwich Enzyme Linked Immunosorbent Assay (s-ELISA). Bacterial pathogens were isolated on blood agar and identified by VITEK&reg; system. 
 
Result: Eleven goats and nine sheep had concurrent infection. Highest positivity for PPRV antigen was observed in nasal and tracheal swab, lungs and lungs associated lymph nodes. Pasteurella multocida was the major bacteria found to cause fibrinious hemorrhagic broncho-alveolar pneumonia followed by Klebsiella pneumoniae spp. pneumoniae producing suppurative bronchoalveolar pneumonia in the presence of PPR infection. Sphingomonas paucimobilis and Acinetobacter baumannii, human pathogen were also isolated and found to produce severe hemorrhagic interstitial pneumonia and serofibrinous bronchopneumonia respectively in presence of PPR which is novel finding. Our study is the first report of necropsy based investigation on concurrent infection in sheep and goats.</div>

Molecular Detection of Some Most Important Pathogenic Bacteria by Multiplex PCR for Diagnosis of Subclinical Mastitis in Bovine

Efficacy of purified antimicrobial peptides from lactic acid bacteria against bovine mastitis pathogens

Evaluation of Efficiency and Extent of Inhibitory Characteristics of Lactobacillus spp. (Lactobacillus plantarum) against Selected Pathogenic Bacteria

Synergetic effect of different free-living diazotrophic bacteria, P – solubilizing bacteria and Rhizobium on growth of Oryza sativa L. (cv. NDR – 359)

Current Status of Nematode-bacteria Interaction: A Mini Review

Environment-Pathogen Interaction in Plant Diseases

Control Activity and Antibiotic Gene Detection of Endophytic Bacteria in Suppressing Cocoa Black Pod Disease (Phytophthora palmivora Butl.)

Potential of Lactic Acid Bacteria in Plant Growth Promotion

Isolation, Molecular Identification and Multidrug Resistance Profiling of Bacteria Causing Clinical Mastitis in Cows

A Necropsy based Investigation on Concurrent Infection of PPR and Emerging Opportunistic Bacterial Pathogens in Sheep and Goat