<div style="text-align: justify;">Brucellosis is the most common zoonotic disease in the Mediterranean region where Turkey is located. Therefore, particularly in this region, it is crucial to follow molecular surveillance methods for the control and eradication of this disease agent, which threatens both human and animal life. We have conducted a molecular-based surveillance study for detecting the origin of the possible reservoir in human brucellosis and transmission of Brucella spp. between humans and ruminants. For the detection of clonal relationships, we isolated Brucella spp. from human and ruminants diagnosed with brucellosis and evaluated them by PFGE and MLVA methods. We found 10 subclusters in PFGE and 22 subclusters in MLVA-16. Human and ruminant isolates were found to co-exist in MLVA-16-m1 and PFGE-a8, a9, a10 subcluster by both methods. In conclusion, the results revealed that these subcluster isolates that co-exist some ruminant and human isolates can be more effective in infecting humans.</div>

Molecular epidemiological characteristics of Brucella spp. isolated from human and animal brucellosis

This review analyses the use of molecular techniques in rumen microbial identification. The use of regular methods like roll tube and most probable number resulted in under estimation of rumen microbial growth. So molecular biology acts as an advanced tool for rumen microbial culture and identification of various new species. These techniques give complete descriptions of individual ruminal populations. Use of molecular techniques like PCR, DDGE and FISH, which also pave pathway for genetic manipulation of rumen microbes in the field of rumen manipulation. So the combination of traditional and molecular assays gives accurate and satisfactory results.

Molecular techniques in rumen biotechnology: A review

Molecular markers are effective tools used to &lsquo;flag&rsquo; the location of a specific gene or the inheritance of a definite trait. Markers are unique DNA fragments that can be identified inside the entire genome. The development of molecular markers combined with high throughput technologies have paved the way for achieving the desirable traits as well as induced biotic and abiotic stress tolerance in plant, which enhanced the crop breeding. Highly polymorphic molecular markers are developed for gene mapping, estimation of genetic diversity, finding out the evolution and phylogeny of crop, analysis of heterosis, assessment of diploid/haploid crops and genotyping of cultivars along with Marker Assisted Breeding (MAB)/Marker Assisted Selection (MAS). These are the most significant objectives for crop breeding. This review reveals about the role of various recently developed molecular markers in the improvement of crop. Molecular markers act as a &ldquo;milestone&rdquo; for the researchers who aim to enhance crop breeding.

Role of Molecular Markers in Crop Breeding: A Review

Background: The porcine sapelovirus (PSV) is a small, non-enveloped, single-stranded, positive-sense, RNA virus of the family Picornaviridae. The PSV infections in pigs have been found associated with diarrhoea, polioencephalomyelitis, pneumonia and reproductive disorders with a high morbidity rate. Despite of its economical importance very few studies are available on the pathology of PSV. The present study was conducted with the aim to investigate the PSV infection and associated pathology in Indian pigs.&nbsp; 
 
Methods: Tissue samples along with intestinal content were collected from a total of 78 necropsied cases for histopathological examination and molecular investigation during April 2019 to August 2020. The amplification of 5&#39; UTR region of PSV was carried out via RT-PCR and confirmed by sequencing. The Genetic characterization of Indian isolate of the PSV was done on the basis of viral 5&#39; UTR gene.&nbsp; 
 
Result: A total of eight out of 78 cases were found positive for the PSV. Catarrhal and haemorrhagic enteritis, thickening and clouding of brain meninges along with congestion of brain and pneumonia was observed as common gross lesions. Microscopic lesions included perivascular cuffing, focal gliosis, neuronophagia, congestion of meningeal and cerebral vessels, interstitial pneumonia, inflammatory changes in the intestinal mucosa and sloughing of villi. The genetic characterization revealed maximum identity of 96.89% with PSV-1 strain PSV-46-V (LC508233) and PSV-1 strain PSV-26-B (LC508232) of Zambia. This study reported the pathological and molecular investigation of PSV from Indian pigs. Further explorative surveillance along with experimental studies in suitable animal model and cell lines are highly warranted for better understanding of PSV pathology in Indian pigs.

Pathology and Molecular Characterization of Porcine Sapelovirus in Indian Pigs

<div style="text-align:justify">Background: The porcine kobuvirus (PKV) is a single-stranded, positive-sense, RNA virus of the family Picornaviridae which composed of small, non-enveloped viruses. The PKV is suspected to cause large outbreaks of diarrhoea and dehydration along with vomition in suckling piglets with a high morbidity rate. Despite of its economical importance very few studies are available on the pathology of PKV. The present study was conducted with the aim to investigate PKV in naturally infected Indian pigs.&nbsp; 
 
Methods: A total of 41 intestinal content and tissue samples from dead pigs were collected from post-mortem facility of, ICAR-IVRI during April, 2019 to July 2020. All the carcasses were subjected to detailed necropsy examination and appropriate tissue samples were collected for histopathological examination and molecular investigation. The amplification of 3D region of PKV was carried out via RT-PCR using the specific primers. The Genetic characterization of Indian isolate of the PKV was done on the basis of viral 3D gene of PKV.&nbsp; 
 
Result: A total of six out of 41 necropsied piglets were found positive for the PKV. The diarrhoea and dehydration were observed as the main clinical signs. The thickening and clouding of brain meninges, congestion of brain, pneumonia along with catarrhal and haemorrhagic enteritis was observed as common gross lesions. Microscopic lesions observed in brain were mild to moderate perivascular cuffing, congestion of meningeal and cerebral vessels, focal gliosis and neuronophagia. Mild to severe interstitial pneumonia and emphysema was noticed in lungs. Intestinal mucosa revealed severe inflammatory changes along with sloughing of villi. The genetic characterization revealed maximum identity of 91-93% with Indian isolate of PKV MH394282 and Chinese strain of PKV FJ459905. This study reported the first pathological investigation of PKV from Indian pigs and described the molecular findings in detail. Further explorative surveillance along with epidemiological studies are highly warranted to find out the real impact of the PKV on Indian pigs.</div>

​Pathological and Molecular Investigation of Porcine Kobuvirus (PKV) from Indian Pigs

<div style="text-align:justify">Rapid industrialization and successful green revolution have introduced a wide array of chemicals into our environment; some of these chemicals entered in ecosystem; gets accumulated and exert serious health and ecological problems. These toxic substances can enter the food chain and emphasize pathological changes which damage either cell, organ or system (circulatory, immune, respiratory, digestive, nervous, reproductive and musculo-skeletal etc.) by altering structure and/or function of biological components; DNA, RNA, proteins, lipids and carbohydrates. This review article provides certain molecular diagnostic techniques used for their robust and accurate detection at molecular level. Investigations conducted during 2020-2021 where various review and research articles were surveyed and then extracted to enlist congestive datum for rapid detection of toxicological changes in animals. In our investigations we concluded that toxic substances present in our environment affects health of animals by altering structure and functioning of biomolecules and their concerned system. These cytological and systemic changes can be detected with the help of molecular diagnostic techniques including dideoxysequencing, pyrosequencing, allele specific RT-PCR, CRISPER/Cas, genotyping and microarrays etc. present collection of data will provide congestive information for rapid toxicological detection at molecular level.</div>

Molecular Diagnostic Strategies for the Detection of Toxicological Changes in Animals: A Review

<div>Background: Group B streptococcus (GBS) or Streptococcus agalactiae is an important pathogen associated with bovine mastitis. The organism is also of public health consequences and may cause variety of infections ranging from neonatal sepsis, pneumonia and meningitis to localized infections and urinary tract infection or arthritisin adult humans. Widespread use of antibiotics in veterinary medicine has led to development of resistance among the pathogens. So there is need for surveillance of antimicrobial resistance to ensure effective treatment. 
 
Methods: Milk samples collected from mastitis affected animals were processed for isolation of&nbsp;Streptococcus agalactiae. The isolates were tested for antimicrobial susceptibility. Molecular characterisation was carried out by PCR to study the occurrence of resistance marker genes and virulence marker genes. RAPD was carried out to study genetic diversity among the isolates. 
 
Result: Six isolates of S. agalactiae were obtained from 182 milk samples. Highest resistance was observed against co-trimoxazole and tetracycline followed by ampicillin. tetM gene and tetO genes could be amplified in four and three isolates, respectively. None of the isolates showed amplification for ermA, ermB, mefA and mefE genes. Three isolates were positive for the five virulence genes tested (glnA, cfb, hylB, scaA and cyl). RAPD analysis demonstrated great intraspecific genetic diversity among the streptococcal isolates.</div>

Isolation, Antibiogram and Molecular Characterization of Group B Streptococci Isolates from Bovine Mastitis

<div>Background: Pathomicrobial and molecular investigations of respiratory system diseases was undertaken on twelve adult buffalo carcasses received for necropsy examination at post mortem facility of Department of Veterinary Pathology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana during the period from August, 2018 to February, 2019.&nbsp; 
 
Methods: After collection of the samples, laboratory work was undertaken in the laboratories of Department of Veterinary Pathology, Department of Animal Biotechnology and Central Laboratory of the College, LUVAS, Hisar, Haryana (India) in the year 2018-2019 regarding the examination of the clinical history, pathological, microbiological and molecular investigations. 
 
Result: The gross pathological changes observed in lungs were variable degree of vascular changes, mild to severe consolidation, fibrin deposition along with adhesions to the thoracic wall. Histopathological findings revealed abnormalities of inflation such as pulmonary emphysema, atelectasis, pulmonary congestion and haemorrhages which was associated with different types of pneumonia viz. fibrinous bronchopneumonia, suppurative giant cell pneumonia, interstitial pneumonia and serous pneumonia. Lung, heart blood and tracheal swab samples collected from the buffalo carcasses revealed eighteen bacterial isolates which were identified by Vitek 2 system. These include E. coli (11 isolates), Salmonella enteric enterica (2 isolates), Acinetobacter ursingii (1 isolate), Staphylococcus haemolyticus (1 isolate), Staphylococcus sciuri (1 isolate), Staphylococcus warneri (1 isolate) and Staphylococcus hominis (1 isolate) mostly belonging to opportunistic pathogen category. E. coli serotypes confirmed from these cases were O83, O149 and O8. The results of in vitro drug sensitivity testing revealed that most of bacterial strains were found sensitive to cefoperazone/sulbactum and co-trimoxazole. Molecular studies confirmed Bovine Herpes Virus-1 (BHV-1) infection in one case with Fibrinous bronchopneumonia through real-time quantitative PCR indicating the prevalence of the infection in the state.</div>

Pathomicrobial and Molecular Investigations of Respiratory System Diseases Affecting Buffaloes

<div>Background: Riemerella anatipestifer (R. anatipestifer) is a gram negative, microaerophilic, non-motile, bipolar bacteria. High genetic diversity and molecular differentiation were reported among field isolates. Although the bacterium causes one of the most economically important duck diseases in the north-eastern region of India, little work has been done on isolation, identification and molecular characterization of the bacteria. Hence, the present investigation was undertaken with a view to characterize the R. anatipestifer isolates from ducks of Assam.</div>

<div> 
Methods: Phenotypic and molecular identification of R. anatipestifer isolates from domesticated ducks of Assam, India were carried out during the period from February, 2019 to January 2020. A total of 624 samples (Ocular swab, throat swab, liver, spleen, kidney, brain, heart, lung) from ducks comprising of apparently healthy, ailing and dead ducks were collected from five districts of Assam, India were processed to isolate and identify the bacteria. The tentative identification of the bacteria was done based on phenotypic characteristics viz., colony morphology, growth characteristics and biochemical reactions. All the phenotypically positive isolates were further subjected to molecular identification based on PCR assay targeting 16S rRNA gene and ERIC sequence. 
 
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<div>Result: The bacteria could be isolated from different field samples. The highest percentage of the samples that yielded the bacteria are from brain (76%) followed by spleen (74%) of dead ducks and less number of ocular swab (33%) from apparently healthy ducks were found positive. Sequencing of the amplified product of the selected R. anatipestifer isolates targeting 16S rRNA gene revealed homology percentage of 96.5-100%. Further, sequences representing five geographical locations were submitted to NCBI gene bank. Phylogenetic studies of the isolates indicated that there is prevalence of at least two genetically different strains of R. anatipestifer in the study area. The study suggested that the R. anatipestifer infection is endemic in Assam causing varying rate of morbidity (39%) and mortality (53%) and molecular based confirmation is necessary besides phenotypic identification.</div>

<div>Isolation and Molecular Characterization of Riemerella anatipestifer from Domesticated Ducks of Assam, India</div>

<div>Background: The estimation of genetic diversity in pigeonpea is crucial for desiging breeding programmes and germplasm conservation. Morphological studies alone do not provide sufficient information to understand genetic diversity. Molecular analysis using SSRs can provide additional information on genetic diversity that can be used for selection of diverse parents. 
 
Methods: The experimental material for present study consisted of 50 diverse genotypes of pigeonpea. The fifty genotypes were planted in Randomized Complete Block Design consisted of three replications during the kharif 2020-21. The estimation of morphological genetic diversity was done by Mahalanobis D2 statistics. The molecular genetic diversity analysis was done by using 30 molecular markers in same genotypes. 
 
Result: The assessment of morphological diversity revealed that the fifty pigeonpea genotypes were grouped into four different clusters with cluster I as the largest cluster (33 genotypes). The molecular markers differentiated the 50 genotypes in five major clusters with cluster II as the largest cluster (24 genotypes). The results of present study suggested that morphological and molecular diversity in pigeonpea is different. On the basis of genetic distance, the genotypes RVSA 2014-1 and PA 406 were found most genetically distant and may be used in hybridization programme to create diverse progenies.</div>