ABSTRACT
Naturally occurring mutations in morphogenetic protein 15 (BMP15) are associated with decreased ovulation rate (OR), litter size (LS), and sterility. It is of a great interest to elucidate BMP15 gene in Cholistani sheep breed to uplift socio-economic status and the knowledge of Cholistani sheep breeding in Southern Punjab, Pakistan. In our study, a total of 50 infertile Cholistani sheep aged between 2-6 years and having no blood relation were screened for BMP15 mutations. For this purpose, a high-quality DNA was extracted from the blood of sheep followed by primer designing, Polymerase Chain Reaction (PCR) amplification, DNA sequencing, and in silico analyses. Out of total 50 samples, 9 samples including case 1 (T3), case 2 (T8), case 3 (T17), case 4 (T22), case 5 (T25), case 6 (T33), case 7 (T40), case 8 (T44), and case 9 (T47) were found positive for a variety of already reported and novel BMP15 mutations. Further in silico analyses of the observed mutations have shown the functional impact of these mutations on different characteristics (molecular weight, theoretical PI, estimated half-life, instability index, sub-cellular localization, and 3D confirmation) of the encoded proteins, possibly altering the normal functionality. In a nutshell, findings of this study have confirmed the possible essential role of the BMP15 mutations in the infertility of the Cholistani sheep.
Mutações de ocorrência natural na proteína morfogenética 15 (BMP15) estão associadas à diminuição da taxa de ovulação (TO), tamanho da ninhada (TN) e esterilidade. Estudar a BMP15 na raça Cholistani para elevar o status socioeconômico e o conhecimento da criação de ovinos Cholistani no sul de Punjab, Paquistão. Em nosso estudo, 50 ovelhas Cholistani inférteis sem parentesco sanguíneo foram rastreadas para mutações BMP15. Para tanto, um DNA de alta qualidade foi extraído do sangue dessas ovelhas, seguido de concepção do primer, amplificação da reação em cadeia da polimerase (PCR), sequenciamento de DNA e análises in silico. Do total de 50 amostras, 9, incluindo caso 1 (T3), caso 2 (T8), caso 3 (T17), caso 4 (T22), caso 5 (T25), caso 6 (T33), caso 7 (T40), caso 8 (T44) e caso 9 (T47), foram consideradas positivas para uma variedade de mutações BMP15 novas e já relatadas. Mais análises in silico das mutações observadas mostraram o impacto funcional dessas mutações em diferentes características (peso molecular, PI teórico, meia-vida estimada, índice de instabilidade, localização subcelular e confirmação 3D) das proteínas codificadas, possivelmente alterando a funcionalidade normal. Nossos achados confirmaram o possível papel essencial das mutações BMP15 na infertilidade de ovelhas Cholistani.
Subject(s)
Animals , Sheep , Infertility , Mutation/geneticsABSTRACT
Abstract Naturally occurring mutations in morphogenetic protein 15 (BMP15) are associated with decreased ovulation rate (OR), litter size (LS), and sterility. It is of a great interest to elucidate BMP15 gene in Cholistani sheep breed to uplift socio-economic status and the knowledge of Cholistani sheep breeding in Southern Punjab, Pakistan. In our study, a total of 50 infertile Cholistani sheep aged between 2-6 years and having no blood relation were screened for BMP15 mutations. For this purpose, a high-quality DNA was extracted from the blood of sheep followed by primer designing, Polymerase Chain Reaction (PCR) amplification, DNA sequencing, and in silico analyses. Out of total 50 samples, 9 samples including case 1 (T3), case 2 (T8), case 3 (T17), case 4 (T22), case 5 (T25), case 6 (T33), case 7 (T40), case 8 (T44), and case 9 (T47) were found positive for a variety of already reported and novel BMP15 mutations. Further in silico analyses of the observed mutations have shown the functional impact of these mutations on different characteristics (molecular weight, theoretical PI, estimated half-life, instability index, sub-cellular localization, and 3D confirmation) of the encoded proteins, possibly altering the normal functionality. In a nutshell, findings of this study have confirmed the possible essential role of the BMP15 mutations in the infertility of the Cholistani sheep.
Resumo Mutações de ocorrência natural na proteína morfogenética 15 (BMP15) estão associadas à diminuição da taxa de ovulação (TO), tamanho da ninhada (TN) e esterilidade. Estudar a BMP15 na raça Cholistani para elevar o status socioeconômico e o conhecimento da criação de ovinos Cholistani no sul de Punjab, Paquistão. Em nosso estudo, 50 ovelhas Cholistani inférteis sem parentesco sanguíneo foram rastreadas para mutações BMP15. Para tanto, um DNA de alta qualidade foi extraído do sangue dessas ovelhas, seguido de concepção do primer, amplificação da reação em cadeia da polimerase (PCR), sequenciamento de DNA e análises in silico. Do total de 50 amostras, 9, incluindo caso 1 (T3), caso 2 (T8), caso 3 (T17), caso 4 (T22), caso 5 (T25), caso 6 (T33), caso 7 (T40), caso 8 (T44) e caso 9 (T47), foram consideradas positivas para uma variedade de mutações BMP15 novas e já relatadas. Mais análises in silico das mutações observadas mostraram o impacto funcional dessas mutações em diferentes características (peso molecular, PI teórico, meia-vida estimada, índice de instabilidade, localização subcelular e confirmação 3D) das proteínas codificadas, possivelmente alterando a funcionalidade normal. Nossos achados confirmaram o possível papel essencial das mutações BMP15 na infertilidade de ovelhas Cholistani.
ABSTRACT
Fecal samples were collected from 120 domestic pigeons to determine the Attributable risk of Capillaria spp. The Capillaria spp. was observed in 64 out of 120 (51%) pigeons (70 males and 50 females) under this study. A total of 64 (39 males and 25 females) were found naturally infected with Capillaria spp. with infection percentage of 51% and 50% in males and females respectively. Qualitative examinations include the direct microscopy and faecal floatation while quantitative examination includes McMaster technique (worms load was calculated per gram of the faeces). Month wise Attributable risk showed that eggs of the worms were found to be abundant in the month of July during the present study (60% to 73%) because of high humidity. Very high and very low temperature is not suitable for the proper development of the eggs. Qualitative and quantitative examination revealed that Capillaria spp. was more prevalent in males (51%) than females (50%) but overall there was no significant difference (P>0.05) in the male and female because both individuals invest equal amount of energy in search of food and incubating the eggs. Different breeds of pigeons gave different Attributable risk in different months during the study. Groups of pigeons from different locations showed different variable Attributable risk. Areas with high humidity were more suitable for the development of eggs, which is the reason why higher Attributable risk was observed in Shahdara (75%) area of Lahore, Pakistan.(AU)
Amostras de fezes foram coletadas de 120 pombos domésticos para determinar os fatores de risco de Capillaria spp. Capillaria spp. foi observado em 64 de 120 (51%) pombos (70 machos e 50 fêmeas) neste estudo. Um total de 64 (39 machos e 25 fêmeas) foram naturalmente infectados com Capillaria spp. sendo 51% em machos e 50% em fêmeas. Exames qualitativos incluem microscopia direta e suspensão de fezes, e exames quantitativos incluem a técnica McMaster (vermes são calculados por grama de fezes). O risco por mês demonstrou que ovos dos vermes foram encontrados em abundância no mês de Julho durante o presente estudo (60% a 73%) por causa da alta umidade. Temperaturas muito altas e muito baixas não são adequadas para o desenvolvimento adequado de ovos. O exame qualitativo e quantitativo revelou que Capillaria spp. era mais prevalente em machos (51%) que em fêmeas (50%), mas no geral não houve diferença significativa (P>0.05) entre machos e fêmeas porque ambos investem a mesma energia na busca por alimento e incubação de ovos. Diferentes raças de pombos tem diferentes riscos em diferentes meses durante o estudo. Grupos de pombos de diferentes locais demonstraram risco diferenciado. Áreas com alta umidade eram mais propensas para o desenvolvimento de ovos, o motivo pelo qual maior risco foi observado em shahdara (75%).(AU)
Subject(s)
Animals , Attributable Risk , Capillaria , Columbidae/parasitology , PakistanABSTRACT
The present study was designed to report the prevalence of Anaplasma sp. in blood samples of Cholistan breed of cattle from Bahawalpur District and to determine the risk factors associated with the prevalence of this parasite. A total of 148 blood samples were randomly collected from apparently healthy cattle. On the sampling sites, data on the characteristics of the animals (species, gender, age) were collected through questionnaires. 47 blood samples (31.8% of total) produced the 577 base pairs DNA fragment specific for 16S rRNA gene of Anaplasma sp. by PCR amplification. Out of 47 Anaplasma sp. positive PCR products, 9 were found to be Anaplasma marginale by restriction with BssNa1 and 9 were confirmed to be Anaplasma phagocytophilum (A. phagocytophilum) as they amplified 550 bp fragment from the amplified MSP 2 gene of this species. Risk factor analysis indicated that the presence of parasite was not limited to a particular sex or age group of the infected animals. Comparison of hematological profile revealed that Anaplasma sp. positive cattle had significantly reduced levels of mean corpuscular volume (P=0.02) and eosinophils (P=0.02) than in parasite negative animals. While studied serum biochemical profile remain unaffected when compared between the two groups.
ABSTRACT
To compare the risk factors for diabetes in children of two different socioeconomic status. A cross-sectional analytical study. Visits to one government [low income] and two private [middle income] schools of Karachi in 1999 and 2000. Patients and A total of 260 children; 157 children [mean age 12.10 +/- 4 years] from low-income group and 103 children [mean age 10.6 +/- 0.9 years] from middle-income group were surveyed. Data of physical fitness score [PFS] and BMI was calculated. Dietary records were taken by 24 hours self-reported diet recall charts of two weekdays. Knowledge about health was obtained by a questionnaire given to children and mothers. A significant difference was found in the mean age of low-income and middle-income groups [p-value < 0.001]. Significant association was seen in low-income group compared to middle-income group on the basis of TV viewing [p-value = 0.04]. BMI [p-value = 0.011] and positive family history of diabetes [p-value < 0.001]. Forty-seven percent [n=74] of children from low-income group while 51% [n=53] of middle-income group had poor knowledge about health. The children in both the groups also consumed inadequate amount of calories, the diet being poor in fruit, vegetables and milk and high in the fat content. Although children in both the groups had increased risk factors for diabetes the difference between the two socioeconomic groups was significant in terms of middle-income children having more risk. Thus, changes in lifestyle and behaviour including diet is needed in this high risk group to prevent future generations from developing diabetes