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Article ; Online: Immunopotential of Pasteurella multocida bivalent outer membrane protein-based vaccine entrapped in aluminum hydroxide nanoparticles.

Pegu, Haladhar / Tamuly, Shantanu / Sharma, Rajeev Kumar / Borah, Probodh / Nath, Rita

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology

2022 Volume 53, Issue 4, Page(s) 2299–2307

Abstract: Swine pasteurellosis is one of the most economically important diseases of pig caused by Pasteurella multocida (P. multocida) capsular types A and D. These organisms are commensals and opportunistic pathogens in the upper respiratory tract in pig. In the ...

Abstract	Swine pasteurellosis is one of the most economically important diseases of pig caused by Pasteurella multocida (P. multocida) capsular types A and D. These organisms are commensals and opportunistic pathogens in the upper respiratory tract in pig. In the present study, we extracted whole outer membrane proteins (OMP) from P. multocida capsular types A and D and were mixed together in the ratio of 1:1 forming bivalent outer-membrane proteins. The bivalent OMP was adsorbed onto aluminum hydroxide nanoparticles. The size of aluminum hydroxide nanoparticles adsorbed outer membrane protein was found to be in the range of 125 to 130 nm. We observed that aluminum hydroxide nanoparticles adjuvanted bivalent OMP-based vaccine elicited quicker immune kinetics in terms of IgG response as compared to aluminum hydroxide microparticles adjuvanted bivalent bacterin vaccine against P. multocida capsular type A and D.
MeSH term(s)	Swine ; Animals ; Pasteurella multocida ; Aluminum Hydroxide ; Vaccines, Combined ; Bacterial Outer Membrane Proteins ; Pasteurella Infections/microbiology ; Nanoparticles
Chemical Substances	Aluminum Hydroxide (5QB0T2IUN0) ; Vaccines, Combined ; Bacterial Outer Membrane Proteins
Language	English
Publishing date	2022-08-03
Publishing country	Brazil
Document type	Journal Article
ZDB-ID	2017175-4
ISSN	1678-4405 ; 1517-8382
ISSN (online)	1678-4405
ISSN	1517-8382
DOI	10.1007/s42770-022-00795-1
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Article ; Online: Calcium phosphate nanoparticles conjugated with outer membrane vesicle of Riemerella anatipestifer for vaccine development in ducklings.

Deka, Naba Jyoti / Kalita, Dhruba Jyoti / Tamuly, Shantanu / Sharma, Rajeev Kumar / Bora, Durlav Prasad / Dutta, Rupam / Hazorika, Mousumi / Chabukdhara, Prasanta / George, Shiney

Microbial pathogenesis

2023 Volume 185, Page(s) 106446

Abstract: Biodegradable calcium phosphate nanoparticles offer a viable substitute for traditional adjuvants such as aluminum in vaccine production. Calcium phosphate nanoparticle adjuvanted with outer membrane vesicle (OMV) of gram negative bacteria may induce ... ...

Abstract	Biodegradable calcium phosphate nanoparticles offer a viable substitute for traditional adjuvants such as aluminum in vaccine production. Calcium phosphate nanoparticle adjuvanted with outer membrane vesicle (OMV) of gram negative bacteria may induce efficient immune response in the host. The present study was carried out to evaluate the potential of a mucosal vaccine formulation of calcium phosphate (CAP) nanoparticle using OMV of Riemerella anatipestifer (RA) as antigen against New Duck disease in ducks. The work was initiated with isolation, identification of RA, followed by OMV production and extraction. The CAP-OMV nanoparticle was prepared and characterized. The efficacy of the vaccine formulation and toxicity were studied in ducks. The average OMV yield in terms of protein concentration was found to be 122.33 ± 3.48 mg per liter of BHI broth. In SDS-PAGE, isolated OMVs exhibited presence of 16 distinct protein bands with molecular weight ranging from 142.1 to 12.1 kDa. Seven protein bands of 74.1, 69.3, 55.5, 50.6, 45.6, 25.1 and 13.1 kDa were detected relatively more distinct. The major bands detected in our findings were 42 kDa, 37 kDa and 16 kDa that corresponds to OmpA, OmpH, P6 respectively. The mean zeta size (±SD) and potential of the nanoparticle were 246.20 ± 0.53 nm and -25.60 ± 5.97 respectively. In transmission electron microscopy (TEM), the nanoparticles exhibited an average diameter of 129.80 ± 11.10 nm and displayed spherical morphology. The median protective dose (PD50) of CAP-OMV nanoparticle was 1881.10 μg of protein. Group I ducks received 3762 μg of protein (entrapped protein in CAP-OMV nanoparticle) via intra nasal route and it showed the highest serum IgG and secretory IgA level than other immunized groups. All experimental ducks were challenged with 10 × LD50 on 35 days of post primary immunization. Group I showed 100 % survivability in the challenge study. No gross and biochemical indication of acute or chronic toxicity were recorded. In conclusion, our results suggest that CAP-OMV nanoparticle can be a safe and efficient mucosal vaccine delivery system for RA, eliciting strong immune response in the host.
MeSH term(s)	Animals ; Ducks/microbiology ; Poultry Diseases/microbiology ; Flavobacteriaceae Infections/prevention & control ; Flavobacteriaceae Infections/veterinary ; Riemerella ; Adjuvants, Immunologic ; Vaccine Development ; Bacterial Vaccines ; Calcium Phosphates
Chemical Substances	Adjuvants, Immunologic ; Bacterial Vaccines ; Calcium Phosphates
Language	English
Publishing date	2023-11-10
Publishing country	England
Document type	Journal Article
ZDB-ID	632772-2
ISSN	1096-1208 ; 0882-4010
ISSN (online)	1096-1208
ISSN	0882-4010
DOI	10.1016/j.micpath.2023.106446
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Article: Effect of dietary supplementation of garlic along with multi-strain probiotic on growth performance, relative organ weights, intestinal morphology, caecal microflora and serum profile of broiler chicken

Shrestha, Reema / Joysowal, Mamata / Saikia, B.N. / Tamuly, Shantanu / Mahanta, Jugadev

Indian journal of animal nutrition. 2020 Dec., v. 37, no. 4

2020

Abstract: The present study was conducted to evaluate the effects of feeding garlic along with multi-strain probiotics on performance of broilers. One hundred eighty no. (N=180) of day old commercial Ven Cobb 50 broiler chicks 45 chicks in each group with 3 ... ...

Abstract	The present study was conducted to evaluate the effects of feeding garlic along with multi-strain probiotics on performance of broilers. One hundred eighty no. (N=180) of day old commercial Ven Cobb 50 broiler chicks 45 chicks in each group with 3 replicates of 15 chicks each by following Randomized Block Design (RBD) and chicks were randomly distributed into 4 treatment groups i.e. T₀, T₁, T₂ and T₃ on the basis of their body weight. Experimental diets were formulated for starter and finisher phases to meet the nutrient requirement as per ICAR (2013) using commonly available ingredient. A corn and soybean meal based diet was used as the control diet (T₀). The experimental diets were formulated by mixing the basal diet with garlic powder (GP) and multi-strain probiotics (MSP) at 0.25, 0.50 or 0.75% (GP+MSP) in diets of groups T₁, T₂ and T₃, respectively. daily weight gain and feed conversion ratio (FCR) was significantly (Pd 0.05) improved in T₂ group as compared to other group. Jejunal villus height and villus width was significantly (P<0.05) higher in T₂ group. The population of cecal microflora was not altered by dietary treatments. Broiler chickens fed the 0.50% (MSP+GP) diet exhibited (P<0.05) lower levels of serum triglycerides and cholesterol compared to control group. Collectively, the results show that dietary supplementation of GP+MSP at 0.50% can be used as a synbiotic for better production and health status in commercial broilers birds.
Keywords	blood serum ; body weight ; broiler chickens ; cecum ; cholesterol ; corn ; dietary supplements ; feed conversion ; garlic ; growth performance ; health status ; ingredients ; jejunum ; probiotics ; soybean meal ; villi ; weight gain
Language	English
Dates of publication	2020-12
Size	p. 377-384.
Publishing place	Animal Nutrition Society Of India
Document type	Article
ZDB-ID	2939390-5
ISSN	2231-6744 ; 0970-3209
ISSN (online)	2231-6744
ISSN	0970-3209
DOI	10.5958/2231-6744.2020.00060.2
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Article: Allelic variability of estrogen receptor (ESR) gene and its effect on litter traits of Doom pigs

Rahman, Mokidur / Phookan, Arundhati / Zaman, Galib Uz / Das, Arpana / Akhtar, Farzin / Tamuly, Shantanu / Choudhury, Hitu / Sarma, Lopa M

Tropical animal health and production. 2021 June, v. 53, no. 2

2021

Abstract: The present study was conducted to identify the polymorphism of estrogen receptor (ESR) gene and its biological association with litter traits (litter size at birth, litter size at weaning, litter weight at birth and litter weight at weaning) of Doom ... ...

Abstract	The present study was conducted to identify the polymorphism of estrogen receptor (ESR) gene and its biological association with litter traits (litter size at birth, litter size at weaning, litter weight at birth and litter weight at weaning) of Doom pigs native to Assam. A total of 50 adult pigs (12 males and 38 females) chosen randomly from three different herds (Herd I, Herd II, and Herd III) were utilized in the present study. Detection of polymorphism of ESR gene was done by means of PCR-RFLP method. The amplified PCR product was digested with Pvu II restriction enzyme. PCR-RFLP analysis of ESR gene revealed polymorphic banding pattern. Two genotypes viz. AA and AB were identified. AA genotype yielded one fragment (120 bp) and AB genotype yielded three fragments (120, 65, and 55 bp). BB genotype was not found in the population under study. The frequencies of A and B alleles were found to be 0.650 and 0.350, respectively, and the genotypic frequencies of ESR gene were found to be 0.300 and 0.700 for AA and AB genotypes, respectively. There was no significant (P > 0.05) effect of ESR genotype on litter traits and the population under study was not in Hardy-Weinberg Equilibrium for ESR gene. Clustal W Multiple alignment of partial sequence of ESR gene revealed single nucleotide changes at 33, 65, 70, 83, and 92ⁿᵈ nucleotide positions. The presence of Pvu II polymorphism and identification of single nucleotide variation of ESR gene opens interesting prospects for improvement of litter traits in Doom pig through selective breeding program, especially based on marker-assisted selection.
Keywords	adults ; animal health ; estrogen receptors ; genetic variation ; genotype ; herds ; litter size ; litter weight ; marker-assisted selection ; restriction endonucleases ; swine ; India
Language	English
Dates of publication	2021-06
Size	p. 316.
Publishing place	Springer Netherlands
Document type	Article
ZDB-ID	603363-5
ISSN	1573-7438 ; 0049-4747
ISSN (online)	1573-7438
ISSN	0049-4747
DOI	10.1007/s11250-021-02756-6
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Article ; Online: RIG-I expression pattern and cytokine profile in indigenous ducks infected with duck plague virus

Chabukdhara, Prasanta / Kalita, Dhruba Jyoti / Tamuly, Shantanu / Bora, Durlav Prasad / Saikia, Deep Prakash / Borah, Sanjib / Hazorika, Mousumi / Borah, Mukul C. / Gogoi, Sophia M. / Deka, Naba Jyoti / Gogoi, Ankita / Bordoloi, Gautam / Khargharia, Sanjib / Pathak, Siddhartha S.

Microbial Pathogenesis. 2023 Aug., v. 181 p.106205-

2023

Abstract: The present study was undertaken to elucidate mRNA expression pattern of RIG-I and serum cytokines profile alterations in indigenous ducks of Assam, India viz. Pati, Nageswari and Cinahanh in response to natural infections of duck plague virus. Field ... ...

Abstract	The present study was undertaken to elucidate mRNA expression pattern of RIG-I and serum cytokines profile alterations in indigenous ducks of Assam, India viz. Pati, Nageswari and Cinahanh in response to natural infections of duck plague virus. Field outbreaks of duck plague virus were attended during the study period for collection of tissue and blood samples. The ducks under study were divided into three distinct groups as per health status i.e. healthy, duck plague infected and recovered. Results from the study revealed that RIG-I gene expression was significantly upregulated in liver, intestine, spleen, brain and PBMC of both infected and recovered ducks. However, fold changes in RIG- I gene expression was lower in recovered ducks as compared to infected ones which indicated continued stimulation of RIG-I gene by the latent viruses. Both serum pro and anti-inflammatory cytokines were elevated in infected ducks as compared to healthy and recovered ducks, indicating activation of inflammatory reactions in the ducks due to virus invasion. The results from the study indicated that innate immune components of the infected ducks were stimulated in order to make an attempt to resist the virus from the infected ducks.
Keywords	Anatid alphaherpesvirus 1 ; blood serum ; brain ; cytokines ; duck plague ; gene expression ; genes ; health status ; intestines ; liver ; pathogenesis ; spleen ; viruses ; India ; Duck plague virus ; RIG-I ; Innate immunity
Language	English
Dates of publication	2023-08
Publishing place	Elsevier Ltd
Document type	Article ; Online
ZDB-ID	632772-2
ISSN	1096-1208 ; 0882-4010
ISSN (online)	1096-1208
ISSN	0882-4010
DOI	10.1016/j.micpath.2023.106205
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Article: Livestock, pets and humans as carriers of methicillin-resistant

Sonowal, Dharitree / Ghatak, Sandeep / Barua, Acheenta Gohain / Kandhan, Srinivas / Hazarika, Razibuddin Ahmed / Sen, Arnab / Das, Samir / Sonowal, Sarat / Sharma, Rajeev Kumar / Tamuly, Shantanu / Phukan, Chimanjita / Sharma, Ajanta / Hussain, Poznur

Veterinary research forum : an international quarterly journal

2023 Volume 14, Issue 7, Page(s) 351–358

Abstract: Staphylococcus ... ...

Abstract	Staphylococcus aureus
Language	English
Publishing date	2023-07-15
Publishing country	Iran
Document type	Journal Article
ISSN	2008-8140
ISSN	2008-8140
DOI	10.30466/vrf.2023.1971891.3672
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Article ; Online: RIG-I expression pattern and cytokine profile in indigenous ducks infected with duck plague virus.

Chabukdhara, Prasanta / Kalita, Dhruba Jyoti / Tamuly, Shantanu / Bora, Durlav Prasad / Saikia, Deep Prakash / Borah, Sanjib / Hazorika, Mousumi / Borah, Mukul C / Gogoi, Sophia M / Deka, Naba Jyoti / Gogoi, Ankita / Bordoloi, Gautam / Khargharia, Sanjib / Pathak, Siddhartha S

Microbial pathogenesis

2023 Volume 181, Page(s) 106205

Abstract	The present study was undertaken to elucidate mRNA expression pattern of RIG-I and serum cytokines profile alterations in indigenous ducks of Assam, India viz. Pati, Nageswari and Cinahanh in response to natural infections of duck plague virus. Field outbreaks of duck plague virus were attended during the study period for collection of tissue and blood samples. The ducks under study were divided into three distinct groups as per health status i.e. healthy, duck plague infected and recovered. Results from the study revealed that RIG-I gene expression was significantly upregulated in liver, intestine, spleen, brain and PBMC of both infected and recovered ducks. However, fold changes in RIG- I gene expression was lower in recovered ducks as compared to infected ones which indicated continued stimulation of RIG-I gene by the latent viruses. Both serum pro and anti-inflammatory cytokines were elevated in infected ducks as compared to healthy and recovered ducks, indicating activation of inflammatory reactions in the ducks due to virus invasion. The results from the study indicated that innate immune components of the infected ducks were stimulated in order to make an attempt to resist the virus from the infected ducks.
MeSH term(s)	Animals ; Ducks ; Immunity, Innate ; Leukocytes, Mononuclear/metabolism ; Cytokines/genetics ; Cytokines/metabolism
Chemical Substances	Cytokines
Language	English
Publishing date	2023-06-18
Publishing country	England
Document type	Journal Article
ZDB-ID	632772-2
ISSN	1096-1208 ; 0882-4010
ISSN (online)	1096-1208
ISSN	0882-4010
DOI	10.1016/j.micpath.2023.106205
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Article ; Online: In Vitro and In Vivo Studies on the Efficacy of Zinc-Oxide and Selenium Nanoparticle in Cryopreserved Goat (Capra hircus) Spermatozoa.

Abedin, Sayed Nabil / Baruah, Anubha / Baruah, Kishore Kumar / Kadirvel, Govindasamy / Katiyar, Rahul / Khargharia, Gautam / Bora, Arundhati / Dutta, Devo Jyoti / Sinha, Sudip / Tamuly, Shantanu / Phookan, Arundhati / Deori, Sourabh

Biological trace element research

2023 Volume 201, Issue 10, Page(s) 4726–4745

Abstract: Different nanoparticles (NPs) are currently being investigated for their potential role as cryoprotectant during semen cryopreservation in several mammalian species. It may be possible to improve semen quality following cryopreservation by ... ...

Abstract	Different nanoparticles (NPs) are currently being investigated for their potential role as cryoprotectant during semen cryopreservation in several mammalian species. It may be possible to improve semen quality following cryopreservation by supplementation of NPs in the freezing extenders. The present study was carried out in semen collected from four (4) Assam Hill Goat bucks (10 ejaculates per buck) to investigate the effect of supplementing zinc oxide (ZnO) and selenium (Se) NPs in Tris-citric acid-fructose yolk (TCFY) extender on in vitro sperm quality and in vivo fertility rate after freeze-thawing. The size morphology and zeta potential of ZnO and Se NPs were evaluated prior to its incorporation in the freezing extender. Qualified semen samples (> 70% progressive motility) were divided into five (5) aliquots and then diluted in TCFY extender containing ZnO and Se NP supplementation at different concentrations (T
MeSH term(s)	Pregnancy ; Animals ; Female ; Male ; Semen/metabolism ; Selenium/pharmacology ; Semen Analysis ; Zinc Oxide/pharmacology ; Goats/metabolism ; Sperm Motility ; Spermatozoa ; Cryopreservation/methods ; Antioxidants/metabolism ; Nanoparticles ; Zinc/pharmacology
Chemical Substances	Selenium (H6241UJ22B) ; Zinc Oxide (SOI2LOH54Z) ; Antioxidants ; Zinc (J41CSQ7QDS)
Language	English
Publishing date	2023-01-04
Publishing country	United States
Document type	Journal Article
ZDB-ID	445336-0
ISSN	1559-0720 ; 0163-4984
ISSN (online)	1559-0720
ISSN	0163-4984
DOI	10.1007/s12011-022-03551-6
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Article ; Online: Association between levels of insulin-like growth factor-1 in serum and seminal plasma with fresh and frozen-thawed semen characteristics in Beetal bucks.

Kumar, Keshav / Dutta, Mitali / Deori, Sourabh / Abedin, Sayed Nabil / Gupta, Mohua Das / Sinha, Sudip / Shome, Arijit / Rongmei, Soihem Diana / Tamuly, Shantanu / Hazarika, Girin / Borah, Pranjal

Reproduction in domestic animals = Zuchthygiene

2023 Volume 59, Issue 1, Page(s) e14499

Abstract: The aim of this study was to investigate the relationship between the levels of insulin-like growth factor-1 (IGF-1) in serum and seminal plasma and the characteristics of semen in Beetal bucks (Capra hircus). A total of 12 adult Beetal bucks were ... ...

Abstract	The aim of this study was to investigate the relationship between the levels of insulin-like growth factor-1 (IGF-1) in serum and seminal plasma and the characteristics of semen in Beetal bucks (Capra hircus). A total of 12 adult Beetal bucks were involved in the study, with each buck providing six ejaculates collected using a standard artificial vagina (n = 72 total). Only qualified semen samples (volume of 0.7 mL, a mass motility rating of 3+ or higher on a 0-+ scale, and individual progressive motility of 80% or more) divided into three fractions were processed for estimation of IGF-1 and other seminal parameters like motility, viability, acrosome integrity, sperm abnormality and superoxide dismutase (SOD) activity. The first and second fraction were diluted and extended with Optixcell extender (1:15 ratio). The first ejaculate fraction was processed for studying fresh semen parameters and the second fraction was cryopreserved for evaluating frozen semen parameters. French mini straws (0.25 mL) were used for semen filling, and polyvinyl alcohol powder of different colours was used for sealing the extended semen. The third fraction of each ejaculate was centrifuged at room temperature (1100 × g for 7 min) to separate the seminal plasma. Additionally, blood samples were taken from each buck on the same day as semen collection, resulting in a total of 36 blood samples. The results revealed a significant positive correlation (r = .4243; p < .05) between the concentration of IGF-1 in both serum and seminal plasma of the Beetal bucks. Furthermore, the concentration of IGF-1 in serum showed significant positive correlations with sperm viability (r = .554; p < .05), acrosome integrity (r = .527; p < .05), post-thaw sperm motility (r = .407; p < .01), post-thaw sperm viability (r = .426; p < .01) and post-thaw acrosome integrity (r = .333; p < .05). However, it had a significant negative correlation with SOD activity in fresh semen (r = -0.458; p < .01). Moreover, the concentration of IGF-1 in seminal plasma demonstrated significant positive correlations with individual progressive motility (r = .341; p < .05), sperm viability (r = .527; p < .05), acrosome integrity (r = .539; p < .05), sperm plasma membrane integrity (r = .464; p < .05), post-thaw sperm motility (r = .644; p < .01), post-thaw sperm viability (r = .643; p < .01), post-thaw acrosome integrity (r = .487; p < .01) and post-thaw sperm plasma membrane integrity (r = .521; p < .01). Additionally, it showed a significant negative correlation with SOD activity in both fresh semen (r = -0.714; p < .01) and frozen semen (r = -0.558; p < .01) of Beetal bucks. Based on these findings, IGF-1 in seminal plasma can be considered as a potential biomarker for the selection of bucks for breeding purposes.
MeSH term(s)	Animals ; Female ; Male ; Cryopreservation/veterinary ; Cryopreservation/methods ; Cryoprotective Agents ; Insulin-Like Growth Factor I ; Insulin-Like Peptides ; Semen ; Semen Analysis/veterinary ; Semen Preservation/veterinary ; Semen Preservation/methods ; Sperm Motility ; Spermatozoa ; Superoxide Dismutase ; Goats/metabolism
Chemical Substances	Cryoprotective Agents ; Insulin-Like Growth Factor I (67763-96-6) ; Insulin-Like Peptides ; Superoxide Dismutase (EC 1.15.1.1)
Language	English
Publishing date	2023-11-02
Publishing country	Germany
Document type	Journal Article
ZDB-ID	1015187-4
ISSN	1439-0531 ; 0936-6768
ISSN (online)	1439-0531
ISSN	0936-6768
DOI	10.1111/rda.14499
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Article ; Online: Allelic variability of estrogen receptor (ESR) gene and its effect on litter traits of Doom pigs.

Rahman, Mokidur / Phookan, Arundhati / Zaman, Galib Uz / Das, Arpana / Akhtar, Farzin / Tamuly, Shantanu / Choudhury, Hitu / Sarma, Lopa M

Tropical animal health and production

2021 Volume 53, Issue 2, Page(s) 316

Abstract	The present study was conducted to identify the polymorphism of estrogen receptor (ESR) gene and its biological association with litter traits (litter size at birth, litter size at weaning, litter weight at birth and litter weight at weaning) of Doom pigs native to Assam. A total of 50 adult pigs (12 males and 38 females) chosen randomly from three different herds (Herd I, Herd II, and Herd III) were utilized in the present study. Detection of polymorphism of ESR gene was done by means of PCR-RFLP method. The amplified PCR product was digested with Pvu II restriction enzyme. PCR-RFLP analysis of ESR gene revealed polymorphic banding pattern. Two genotypes viz. AA and AB were identified. AA genotype yielded one fragment (120 bp) and AB genotype yielded three fragments (120, 65, and 55 bp). BB genotype was not found in the population under study. The frequencies of A and B alleles were found to be 0.650 and 0.350, respectively, and the genotypic frequencies of ESR gene were found to be 0.300 and 0.700 for AA and AB genotypes, respectively. There was no significant (P > 0.05) effect of ESR genotype on litter traits and the population under study was not in Hardy-Weinberg Equilibrium for ESR gene. Clustal W Multiple alignment of partial sequence of ESR gene revealed single nucleotide changes at 33, 65, 70, 83, and 92
MeSH term(s)	Alleles ; Animals ; Female ; Gene Frequency ; Genotype ; Litter Size/genetics ; Male ; Polymorphism, Restriction Fragment Length ; Pregnancy ; Receptors, Estrogen/genetics ; Sus scrofa ; Swine/genetics
Chemical Substances	Receptors, Estrogen
Language	English
Publishing date	2021-05-12
Publishing country	United States
Document type	Journal Article
ZDB-ID	603363-5
ISSN	1573-7438 ; 0049-4747
ISSN (online)	1573-7438
ISSN	0049-4747
DOI	10.1007/s11250-021-02756-6
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