Pathology of Pullorum Disease and Molecular Characterization of Its Pthogen

2020 ◽  
Vol 23 (1) ◽  
pp. 25-35
Author(s):  
MM Rahman ◽  
J Alam ◽  
MM Rahman ◽  
MAHNA Khan ◽  
MG Haider
Keyword(s):  
Molecular Characterization ◽  
Molecular Identification ◽  
Intestinal Mucosa ◽  
Inflammatory Cells ◽  
Liver Parenchyma ◽  
Histopathological Study ◽  
Isolation And Identification ◽  
Focal Necrosis ◽  
Salmonella Pullorum

This experiment was conducted to determine the pathology of pullorum disease of chickens and molecular identification of its pathogen. A total of 108 samples, including swabs from different organs were collected from different commercial layer farms of Gazipur district. The histopathological samples were collected in 10% formalin and the swabs were collected in tetrathionate broth. Samples were subjected to isolation and identification of the causal agent followed by gross and histopathological study of the affected visceral organs. Fifty eight out of 108 cloacal swabs (53.7%) were Salmonella positive. The percentage of Salmonella positive in liver swabs, spleen swabs, lung swabs and intestinal swabs from dead birds were 55.88%, 32.35%, 35.29% and 47.05%, respectively. On average, 52.94% livers of Salmonella affected birds were enlarged, congested and hemorrhagic and necrotic foci was present in 32.35% liver. Unabsorbed and coagulated yolk was found in 70.58% cases. From these 38.24% spleens were swollen and congested and 44.12 % kidneys were enlarged. At histopathology, 52.94% livers showed congestion, focal necrosis with multifocal infiltration of histiocytes in liver parenchyma. Focal necrosis and inflammatory cells were found in 70.58% spleen. Infiltration of heterophils in intestinal mucosa was found in 47.05% cases. 20.58% (7 out of 34) samples were PCR positive for Salmonella Pullorum organism. Ann. Bangladesh Agric. (2019) 23(1) : 25-35

Incidence and molecular characterization of fungi and yeast isolated from cultured catfish and Nile tilapia

2020 ◽  
Vol 21 (3) ◽  
pp. 61-66
Author(s):  
Ola Hashem ◽  
Viola Zaki ◽  
Rawia Adawy
Keyword(s):  
Molecular Characterization ◽  
Oreochromis Niloticus ◽  
Nile Tilapia ◽  
Clarias Gariepinus ◽  
Fungal Species ◽  
High Rate ◽  
Fish Farms ◽  
Isolation And Identification ◽  
Penicillium Spp

Objective: To study the incidence and seasonal dynamics of different fungi affected freshwater fishes in Lake Manzala with molecular identification of the isolated fungi. Animals: 300 Nile tilapia (Oreochromis niloticus) and 300 catfish (Clarias gariepinus). Design: Descriptive study. Procedures: Random samples of Oreochromis niloticus (O. niloticus) and Clarias gariepinus (C. gariepinus) were collected from Manzala fish farms. Clinical and postmortem examination of fish was applied. Isolation and identification of different fungi were performed by conventional methods. Furthermore, the molecular characterization of isolated fungi was carried out. Results: C. gariepinus had a higher rate of infection with different fungal species than O. niloticus. Aspergillus spp. (Aspergillus niger and Aspergillus flavus) were the most fungal isolated from the examined fishes, followed by Penicillium spp. and Candida albicans. Aspergillus spp were detected in all seasons with a higher rate in summer and spring. A. flavus, A. niger, Penicillium spp. and C.albicans isolates were amplified from both C. gariepinus and O. niloticus at the specified molecular weight using PCR. Conclusion and clinical relevance: Fungal infection affected the fish showing different external and internal lesions, all species of Aspergillus were found in all seasons with a high rate in, hot seasons, summer and spring. The Prevalence of Penicillium and C. albicans were also reported. All fungal isolates were identified on the phenotypic and molecular bases.

Molecular Detection and Phylogenetic Analysis of Chicken Astrovirus Associated with Poultry Enteritis

2021 ◽  
Author(s):  
C. Patidar ◽  
D.K. Sharma ◽  
R. Singathia ◽  
P. Suthar ◽  
A. Saraswat ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Molecular Characterization ◽  
Histopathological Examination ◽  
Villous Atrophy ◽  
Inflammatory Cells ◽  
Liver Parenchyma ◽  
Amino Acid Sequences ◽  
Poultry Birds ◽  
Commercial Poultry ◽  
Liver Specimen

Background: Poultry enteritis is an important multifactorial disease. Chicken Astrovirus (CAstV) usually associated with enteritis. The aim of this study was to investigate the occurrence of CAstV in poultry enteritis cases, its molecular characterization, phylogenetic analysis and gross and microscopic examination of intestine and liver specimen affected with CAstV. Methods: Total 604 dead poultry birds from commercial poultry farms affected with enteritis were examined for presence of CAstV. Intestinal samples of four birds were pooled to make one biological sample. CAstV was detected by reverse transcriptase PCR (RT-PCR) using ORF-1b gene specific primers. Molecular characterization was carried out by partial gene sequencing. Result: CAstV was detected in 20.52% (31/151) of samples. Highest prevalence (49.29%) was observed in 0-1 week old chicks. The partial molecular characterization revealed high similarity of the nucleotide sequence from India (97% to 93%) and from USA, Brazil, Poland and Korea (94 to 92%). Further similarity of amino acid sequences of CAstV from India (100% to 98%) and from USA, Brazil, Poland and Korea (98 to 97%) was observed. Histopathological examination revealed villous atrophy, congestion and atrophic cystic glands in sub-mucosa of intestine. Further severe congestion and hemorrhages along with infiltration of inflammatory cells in liver parenchyma was observed.

scholarly journals Seroprevalence and pathological study of Salmonella infections in layer chickens and isolation and identification of causal agents

1970 ◽  
Vol 4 (2) ◽  
pp. 79-85 ◽  
Author(s):  
MM Islam ◽  
MG Haider ◽  
EH Chowdhury ◽  
M Kamruzzaman ◽  
MM Hossain
Keyword(s):  
Mononuclear Cells ◽  
Research Work ◽  
Liver Parenchyma ◽  
Muscularis Mucosa ◽  
Pathological Study ◽  
Isolation And Identification ◽  
Bacterial Colony ◽  
Focal Necrosis ◽  
Salmonella Infections ◽  
Layer Chickens

The present research work was conducted to study about the seroprevalence and pathology of Salmonella infections in layer chickens of Dhaka and Gazipur regions of Bangladesh and to isolate and characterize Salmonellae from layer chickens during the period from January to May 2006. The used materials were blood sample, cloacal and liver swabs from live and dead birds respectively, and visceral organs (liver, lungs, spleen and intestine). The used methods were serum plate agglutination (SPA) test; necropsy and histopathology; cultural, morphological and biochemical test. The overall seroprevalence was 43.4%. During necropsy, congested and enlarged liver with focal necrosis, haemorrhagic and discoloured ovary with stalk formation and mild haemorrhagic to catarrhal enteritis in intestine were detected. Microscopically, the liver showed congestion, focal necrosis with multifocal infiltration of histiocytes in liver parenchyma. In ovum, infiltration of leukocytes, bacterial colony, RE cell proliferation and fibrinoid necrosis were recorded. The intestinal mucosa exhibited infiltration of mononuclear cells in mucosa, submucosa as well as muscularis mucosa. A total of 33 (21.02%) Salmonellae from live and dead birds were isolated. The isolation rate of Salmonellae was higher in seronegative (31.6%) group than seropositive (3.2%) group. Out of 33 Salmonella isolates, 25 were S. pullorum, 3 were S. gallinarum and the rest 5 were motile Salmonellae. The isolates obtained in the study may be investigated for serotyping, pathogenesis study, antibiogram and vaccine production in future. Key words: Seroprevalence, histopathology, isolation, identification, Salmonella, chickensDOI = 10.3329/bjvm.v4i2.1288Bangl. J. Vet. Med. (2006). 4 (2): 79–85

scholarly journals Seroprevalence and pathology of naturally infected Salmonellosis in poultry with isolation and identification of causal agents

1970 ◽  
Vol 6 (2) ◽  
pp. 327-334 ◽  
Author(s):  
AKM Ahmed ◽  
MT Islam ◽  
MG Haider ◽  
MM Hossain
Keyword(s):  
Antibiotic Sensitivity ◽  
Inflammatory Cells ◽  
Agglutination Test ◽  
Isolation And Identification ◽  
Salmonella Gallinarum ◽  
Dead Bird ◽  
Focal Necrosis ◽  
Salmonella Infections ◽  
Leukocytic Infiltration ◽  
Egg Follicles

The present study was conducted to determine the seroprevalence of Salmonella infections in poultry. This study covered cultural prevalence with isolation and identification of the causal agent and pathological lesions in different organs produced by Salmonella in layer farms of Mymensingh district during July-December 2007. The materials were blood samples, liver and cloacal swabs from live and dead birds. The used methods were whole blood agglutination test by commercially available Salmonella antigen kit, culture in different media, Gram's staining, motility test, basic 5 sugar fermentation test and histopathology. The overall seroprevalence was 45.9% in live bird and the rate of seroprevalence decreased with advancement of age of birds. The cultural prevalence in seropositive group was 71% and in seronegative group was 59%. In dead bird, the cultural prevalence in liver was 64% and from cloaca 57%. A total 160 isolates were characterized, among them 64.2% were Salmonella Pullorum, 22.3% were Salmonella Gallinarum and 13.5% were Paratyphoid group of Salmonellae. Pathologically friable, bronze color liver with focal necrosis, various grades of enteritis, and hemorrhagic and congested egg follicles with stalk formation were found. Microscopically, focal necrosis and degeneration with leukocytic infiltration in liver, inflammatory cells in the mucosa and submucosa of intestine were found. In egg follicles, congestion and hemorrhages with leukocytic infiltration were found. The isolated Salmonella organisms may be used for further research such as serotyping, vaccine production and antibiotic sensitivity test. Keywords: Salmonellosis; Seroprevalence; Agglutination test; Histopathology; Poultry DOI: 10.3329/jbau.v6i2.4830 J. Bangladesh Agril. Univ. 6(2): 327-334, 2008

Isolation and Molecular Characterization of Riemerella anatipestifer from Domesticated Ducks of Assam, India

2021 ◽  
Author(s):  
M.K. Doley ◽  
S. Das ◽  
R.K. Sharma ◽  
P. Borah ◽  
D.K. Sarma ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Molecular Characterization ◽  
Molecular Identification ◽  
Rrna Gene ◽  
Eastern Region ◽  
Riemerella Anatipestifer ◽  
Field Samples ◽  
Apparently Healthy

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.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.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.

scholarly journals Evaluation of antibacterial activity and molecular characterization of bacteria from Holothuria atra intestine collected from anthropogenic and non-anthropogenic region in Karimunjawa, Indonesia

2020 ◽  
Vol 21 (7) ◽  
Author(s):  
Bambang Sulardiono ◽  
NINIEK WIDYORINI ◽  
DJOKO SUPRAPTO ◽  
DIAH AYUNINGRUM ◽  
ARIF RAHMAN
Keyword(s):  
Antibacterial Activity ◽  
Molecular Characterization ◽  
Molecular Identification ◽  
Sea Cucumber ◽  
Bacterial Isolates ◽  
Plate Method ◽  
Agar Plug ◽  
16S Rdna Pcr ◽  
Holothuria Atra

Abstract. Sulardiono B, Widyorini N, Suprapto D, Ayuningrum D, Rahman A. 2020. Evaluation of antibacterial activity and molecular characterization of bacteria from Holothuria atra intestine collected from anthropogenic and non-anthropogenic region in Karimunjawa, Indonesia. Biodiversitas 21: 3149-3155. Evaluation of antibacterial activity and molecular characterization of bacteria from the intestine of Holothuria atra is needed in the anthropogenic region of Menjangan Besar, Karimunjawa. The research aims to evaluate antibacterial activity and molecular characterization of bacteria from intestine of Holothuria atra collected from anthropogenic and non-anthropogenic regions in Karimunjawa, Indonesia. Sea cucumber samples were collected at Menjangan Besar waters as anthropogenic region (code of HM) and Alang-alang waters as non-anthropogenics region (code of HA), Karimunjawa National Park, Indonesia. The H. atra sample collection was using purposive sampling method. Examination of the bacteria from isolation until molecular characterisation was done at the Tropical Marine Biotechnology Laboratory, Uiversitas Diponegoro. The isolation process was conducted using spread plate method, followed by bacterial isolates purification using streak plate method. Screening of antibacterial activity using the agar plug method, as well as molecular identification was conducted by 16S rDNA PCR amplification. The results of this study indicated that a total of 26 bacterial strains were successfully isolated from sea cucumber intestine, consisted of eleven bacterial isolates from H. atra in Menjangan Besar waters and the rest fifteen isolates from H. atra in Alang-alang waters. The preliminary assay of antibacterial test showed H. atra in both waters have potential bacterial isolates. Those bacterial isolates with antibacterial activity were HM1.2 and HA1.1, which based on molecular identification, showed isolates HM1.2 and HA1.1 had the closest similarity with Bacillus paramcoides and Vibrio alginolyticus consecutively, with BLAST homology 98% and 99%. The accession number for both isolates were LC550090 and LC550089, respectively.

scholarly journals Histopathological study of the Influence of Estradiol on Liver Regeneration in Male Rabbit

2013 ◽  
Vol 37 (2) ◽  
pp. 211-217
Author(s):  
Mahdi A. K.
Keyword(s):  
Liver Regeneration ◽  
Intramuscular Injection ◽  
Inflammatory Cells ◽  
Liver Parenchyma ◽  
Necrotic Area ◽  
Control Group ◽  
Histopathological Study ◽  
Central Vein ◽  
Generative Capacity ◽  
Post Operation

This study was investigated to recognize the generative capacity of liver that influenced by administration of estradiole benzoate. Twenty four healthy local breed male rabbits weighed 1.5-2 kg were used .They were classified into two equal groups control and treated groups. Animals anaesthetized by intramuscular injection of Ketamine (35mg /kg B.W) and Xylazine (5mg\kg B.W.) In control group partial hepatectomy of liver was done, while in treated animals were subject to a same operation then received an intramuscular injection 0.1ml of estradiol benzoate 0.2% at 3rd and 5th days post operation. At 3rd days microscopic finding showed biopsy were taken at 3rd, 7th and 14 day, post operation to study the histopathological changes in liver regeneration .At 3rd day it showed early proliferation of hepatocyte character by irregular hyperchromatic multinucleated cell .While in control group it showed necrotic area of liver parenchyma surrounded by thick fibrous connective tissue infiltrated with inflammatory cells. At 7th day in treated animals still progress in proliferation of hepatocyte while, liver proliferation phase showed in control group .At 14th day in treated animals showed large cells of hepatocyte with double nuclei aggregation without central vein , while in control group showed small hyperchromatic hepatocyte with present of necrotic area. In conclusion the results of this study indicate that administration of estrogen after partial resection in male rabbits has a role in liver regeneration by early initiate or facilitate heypatocyte proliferation.

scholarly journals Pathogenesis of Pullorum Disease (PD) in Chickens by Local Isolate of Salmonella pullorum in Bangladesh

2014 ◽  
Vol 11 (2) ◽  
pp. 1-16
Author(s):  
MG Haider ◽  
EH Chowdhury ◽  
SMK Sharif ◽  
MM Hossain
Keyword(s):  
Egg Production ◽  
Clinical Signs ◽  
Inflammatory Cells ◽  
Experimental Period ◽  
Reproductive Organs ◽  
Control Group ◽  
Histopathological Study ◽  
Nodule Formation ◽  
Time Interval ◽  
Salmonella Pullorum

Pullorum disease is caused by Salmonella enterica subspecies enterica serovar Pullorum. Chickens are the natural host of this pathogen. In the present study experimental pathogenesis was studied. Twenty pullets (Salmonella pullorum seronegative) of Isa Brown breed of 18 weeks and 10 cocks (Salmonella pullorum seronegative) of RIR breed of 26 weeks of age were experimentally infected orally with 2 x107 (CFU) dose of Salmonella Pullorum organisms and in control group no bacteria was given. Birds were observed for clinical signs, gross pathology, and reisolation of S. Pullorum from different organs and blood, histopathological study, detection of antibody levels and detection of S. Pullorum by PCR at different time intervals of experimental period. Four hens and one cock were randomly selected and sacrificed on 6 hr before inoculation and 1 wk, 2, 3 and 4 wks of post infection (PI). Samples were collected for bacteriological, serology and histopathological examinations. Liver, lungs, ovarian follicles and testis were also collected in 50% buffered- glycerol and preserved in - 80&edeg;C for PCR. The clinical signs of infected hens were found at 72 hrs of PI, which continued up to 4 wks. 15.81% reduction in egg production was observed. The highest mean CFU ml-1 of Salmonella Pullorum from blood was 13.55x 103 at 1 wk PI and the lowest was 13x 102 at 4 wk PI. Gross lesions were variable in different birds at different time interval. The highest gross lesion was 93.75% as swollen and congested spleen and the lowest lesion was 43.75% as pericarditis and necrotic foci/ nodules in the heart. Microscopically, the liver showed congestion; hepatitis with infiltration of inflammatory cells, and focal necrosis with nodule formation. The antibody titre increased gradually and the highest titer was at 4 wks PI in hens (4712±1851) than that of cock (3059±903). S. Pullorum was detected by PCR in all liver and lung samples from 1 wk to 4 wks PI. S. Pullorum was reisolated from male and female reproductive organs after experimental infection. S. Pullorum was detected by PCR at 1 wk to 3 wks PI from testicular tissues. S. Pullorum was also reisolated from 50% eggs of experimentally infected birds. DOI: http://dx.doi.org/10.3329/sja.v11i2.18398 SAARC J. Agri., 11(2): 1-16 (2013)

scholarly journals Salmonellosis in layer chickens: pathological features and isolation of bacteria from ovaries and inner content of laid eggs

2012 ◽  
Vol 10 (1) ◽  
pp. 61-67 ◽  
Author(s):  
A K Saha ◽  
M A Sufian ◽  
M I Hossain ◽  
M M Hossain
Keyword(s):  
Dead Layer ◽  
Histopathological Study ◽  
Pathological Study ◽  
Isolation Rate ◽  
Isolation And Identification ◽  
Salmonella Gallinarum ◽  
Visceral Organs ◽  
Leukocytic Infiltration ◽  
Salmonella Pullorum ◽  
Egg Follicles

We isolated and identified Salmonella organisms from ovaries of dead layer birds and from inner content of laid eggs of different poultry farms. The thirty eight ovarian swabs for bacteriology, visceral organs (liver, lung, spleen, egg follicles and intestine) of 38 dead birds for pathological study from 15 layer farms and 45 laid eggs (5 eggs/farm) from reported 9 Salmonella infected farms constituted samples of the study. Samples were subjected to isolation and identification of the causal agent followed by gross and histopathological study of the affected visceral organs. Seventeen of 38 ovarian swabs (44.7%) were Salmonella positive. The percentage of Salmonella Gallinarum, Salmonella Pullorum and paratyphoid causing Salmonella were 70.6%, 17.6% & 11.8%, respectively. About 53% livers of Salmonella affected birds were enlarged, congested, friable and bronze coloured with white necrotic foci. About 59% egg follicles were congested, hemorrhagic, discoloured with stalk formation and 70.6% intestines showed hemorrhagic to catarrhal enteritis. At histopathology, 76.5% livers were congested with formation of multifocal nodules and 82.4% egg follicles were congested with huge leukocytic infiltration. Infiltration of heterophils in intestinal mucosa was found in 47.1% cases. Four Salmonella isolates were found from 9 laid egg samples (5 eggs content comprised as 1 sample) and isolation rate was 44.4% reporting transovarian transmission in poultry Salmonellosis.   DOI: http://dx.doi.org/10.3329/jbau.v10i1.12095   J. Bangladesh Agril. Univ. 10(1): 61–67, 2012  

scholarly journals Molecular characterization of two AP2/ERF transcription factor genes from Egyptian tomato cultivar (Edkawy)

2017 ◽  
Vol 4 (1) ◽  
pp. 12 ◽  
Author(s):  
Kareem Mosa ◽  
Eslam Heb El-din ◽  
Ahmed Ismail ◽  
Fawzy El- Feky ◽  
Ali El-Refy
Keyword(s):  
Transcription Factor ◽  
Abiotic Stress ◽  
Drought Stress ◽  
Stress Tolerance ◽  
Molecular Characterization ◽  
Abiotic Stress Tolerance ◽  
Isolation And Identification ◽  
Tomato Cultivar ◽  
Erf Transcription Factor

The tomato is ranked first amongst vegetable crops in Egypt in relation to surface area and production. The Egyptian tomato cultivar Edkawy has shown abiotic stress tolerance characteristics. However, there is not much information about the molecular characterization of this cultivar. Furthermore, information regarding the identification of abiotic stress tolerance genes from the Edkawy tomato cultivar is lacking. Here, we investigated the ability of the Edkawy cultivar to tolerate drought stress. Two varieties were used as a control in this study; Peto86 (sensitive variety) and Strain B (tolerant variety). Edkawy, Peto86 and Strain B varieties were exposed to drought stress by reducing the water supply gradually. Interestingly, Edkawy demonstrated a remarkable tolerance phenotype to drought stress. Furthermore, we identified and isolated two members of the AP2/ERF transcription factor family from Edkawy which are associated with abiotic stress, particularly drought, i.e. ERF1 and ERF5. Protein prediction, validation and active site prediction of ERF1 and ERF5 were also determined. In addition to the domain obtained by the pfam online tool, the interaction between Edkawy ERFs proteins and other proteins in the Solanaceae family was obtained. Furthermore, subcellular localization was determined by the ngLOC and Plant-mPLoc online tools. Characterization of the Edkawy tomato cultivar and isolation and identification of such transcription factors will help in the engineering of tomato plants with abiotic stress tolerance.

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