Enterotoxigenic methicillin resistant Staphylococcus aureus contamination in salted fish from Gwadar Balochistan / Contaminação de Staphylococcus aureus resistente à meticilina enterotoxigênica em peixes salgados de Gwadar Balochistan

Staphylococcus aureus is an important foodborne pathogen associated to food intoxication and other multiple infections in human being. Its presence in salted food is a serious issue due to its salt tolerance potential. A study was conducted to analyze the presence of enterotoxins producing drug resistance S. aureus in salted sea fish from Gwadar. Freshly persevered samples (n=50) of salted fish were subjected to analyze the presence of S. aureus using 16S rRNA and Nuc genes primers. The isolates were then evaluated for drug resistance and enterotoxins producing potential using specific primers for MecA (methicillin resistance gene), (SEA) staphylococcal enterotoxin A and (SEB) staphylococcal enterotoxin B genes. Total 13/50 (26%) of the samples were found positive for the presence of S. aureus, preliminary confirmed with biochemical profiling and finally with the help of target genes presence. The isolates were found showing 100% resistant to methicillin, which were molecularly confirmed by the presence of MecA gene present in genome. The isolates 5/13 (38%) were positive for SEA and 3/13 (23%) for SEB genes, whereas 2/13 (15%) were confirmed having both SEA and SEB genes in its genome. It was also confirmed that all the isolates were capable to form biofilm over the glass surfaces. It was concluded that the study confirmed the presence of enterotoxigenic methicillin resistance Staphylococcus aurous (MRSA) in salted fish product, that poses gross food safety concern. Preventive and control measures are necessary to handle this serious food safety concern.

Staphylococcus aureus é um importante patógeno de origem alimentar associado à intoxicação alimentar e outras infecções múltiplas em seres humanos. Sua presença em alimentos salgados é um problema sério devido ao seu potencial de tolerância ao sal. Um estudo foi realizado para analisar a presença de enterotoxinas produtoras de resistência a drogas S. aureus em peixes salgados do mar de Gwadar. Amostras recém-perseveradas (n = 50) de peixes salgados foram submetidas à análise da presença de S. aureus usando os primers dos genes 16S rRNA e Nuc. Os isolados foram então avaliados quanto à resistência a drogas e potencial de produção de enterotoxinas usando primers específicos para os genes MecA (gene de resistência à meticilina), (SEA) enterotoxina A estafilocócica e (SEB) enterotoxina B estafilocócica genes. Um total de 13/50 (26%) das amostras foi considerado positivas para a presença de S. aureus, confirmadas preliminarmente com perfis bioquímicos e finalmente com a ajuda da presença de genes-alvo. Os isolados foram encontrados com 100% de resistência à meticilina, os quais foram confirmados molecularmente pela presença do gene MecA no genoma. Os isolados 5/13 (38%) foram positivos para SEA e 3/13 (23%) para genes SEB, enquanto 2/13 (15%) foram confirmados tendo os genes SEA e SEB em seu genoma. Também foi verificado que todos os isolados foram capazes de formar biofilme sobre as superfícies de vidro. Concluiu-se que o estudo confirmou a presença de Staphylococcus aurous resistente à meticilina enterotoxigênica (MRSA) em produtos de peixe salgado, o que representa uma grande preocupação para a segurança alimentar. Medidas preventivas e de controle são necessárias para lidar com essa grave preocupação com a segurança alimentar.

Enterotoxigenic methicillin resistant Staphylococcus aureus contamination in salted fish from Gwadar Balochistan

Abstract Staphylococcus aureus is an important foodborne pathogen associated to food intoxication and other multiple infections in human being. Its presence in salted food is a serious issue due to its salt tolerance potential. A study was conducted to analyze the presence of enterotoxins producing drug resistance S. aureus in salted sea fish from Gwadar. Freshly persevered samples (n=50) of salted fish were subjected to analyze the presence of S. aureus using 16S rRNA and Nuc genes primers. The isolates were then evaluated for drug resistance and enterotoxins producing potential using specific primers for MecA (methicillin resistance gene), (SEA) staphylococcal enterotoxin A and (SEB) staphylococcal enterotoxin B genes. Total 13/50 (26%) of the samples were found positive for the presence of S. aureus, preliminary confirmed with biochemical profiling and finally with the help of target genes presence. The isolates were found showing 100% resistant to methicillin, which were molecularly confirmed by the presence of MecA gene present in genome. The isolates 5/13 (38%) were positive for SEA and 3/13 (23%) for SEB genes, whereas 2/13 (15%) were confirmed having both SEA and SEB genes in its genome. It was also confirmed that all the isolates were capable to form biofilm over the glass surfaces. It was concluded that the study confirmed the presence of enterotoxigenic methicillin resistance Staphylococcus aurous (MRSA) in salted fish product, that poses gross food safety concern. Preventive and control measures are necessary to handle this serious food safety concern.

Resumo Staphylococcus aureus é um importante patógeno de origem alimentar associado à intoxicação alimentar e outras infecções múltiplas em seres humanos. Sua presença em alimentos salgados é um problema sério devido ao seu potencial de tolerância ao sal. Um estudo foi realizado para analisar a presença de enterotoxinas produtoras de resistência a drogas S. aureus em peixes salgados do mar de Gwadar. Amostras recém-perseveradas (n = 50) de peixes salgados foram submetidas à análise da presença de S. aureus usando os primers dos genes 16S rRNA e Nuc. Os isolados foram então avaliados quanto à resistência a drogas e potencial de produção de enterotoxinas usando primers específicos para os genes MecA (gene de resistência à meticilina), (SEA) enterotoxina A estafilocócica e (SEB) enterotoxina B estafilocócica genes. Um total de 13/50 (26%) das amostras foi considerado positivas para a presença de S. aureus, confirmadas preliminarmente com perfis bioquímicos e finalmente com a ajuda da presença de genes-alvo. Os isolados foram encontrados com 100% de resistência à meticilina, os quais foram confirmados molecularmente pela presença do gene MecA no genoma. Os isolados 5/13 (38%) foram positivos para SEA e 3/13 (23%) para genes SEB, enquanto 2/13 (15%) foram confirmados tendo os genes SEA e SEB em seu genoma. Também foi verificado que todos os isolados foram capazes de formar biofilme sobre as superfícies de vidro. Concluiu-se que o estudo confirmou a presença de Staphylococcus aurous resistente à meticilina enterotoxigênica (MRSA) em produtos de peixe salgado, o que representa uma grande preocupação para a segurança alimentar. Medidas preventivas e de controle são necessárias para lidar com essa grave preocupação com a segurança alimentar.

Characterization of laser-induced shock waves generated during infrared laser ablation of copper by the optical beam deflection method.

The shock waves generated during laser ablation of a copper target are investigated using the optical beam deflection method. The fluence of nanosecond pulsed infrared laser beam was in the range of 15-700J/cm2. The density jumps related with the influx of the shock wave at two interaction points were detected with the help of He-Ne laser probes. In general, a supersonic shock wave is produced, which propagates through air and gradually decays into an acoustic wave. Experiments were carried out to study the impact of laser fluence and propagation distance on the shock wave velocity and pressure. The shock wave velocity varies with laser fluence as v∝Fl0.3 and with propagation distance as v∝d-1.5. These results are compared with the predictions of the theoretical models. In the investigated fluence range, shock wave pressure rises by an order of magnitude (∼1-10MPa). We demonstrated that shock wave pressure and ablated mass can be related, yielding mass-specific shock wave pressure that increases linearly with laser fluence. We have also noticed the shock-wave-induced probe beam focusing under certain conditions, which indicates that the shock wave modifies the refractive index of the compressed layer of air. The reported results are useful for the fundamental understanding and pave the way for new applications of laser-induced shock waves.

Legal framework in aid of biological diversity and statutory deficiencies in Pakistan.

The traditional perception of mutual rights and obligations in the developing world was practically confined to human beings only. Fortunately, this conventional understanding is growing to include all living beings in the scope of certain basic rights. It is also addressing those creatures which are co-existing with human societies naturally or are associated for their subsistence and interdependence. It means that there should be prescribed body of rules and regulations to regulate human conduct in the society. In Pakistan social awakening for animal rights supported by international campaigns have urged state institutions to make adequate body of rules and regulations for protection of animal rights. Purpose of developing required understanding, at the first place is to clarify what is a right and what is an obligation? Secondly what should be the nature of these rights and obligations. Should these be only specific to human beings or there are other beings who also possess certain rights? If other living beings, species have rights, how these rights are to be conferred and regulated? Who is under obligation to provide these rights? Who is liable and responsible to ensure observance of such obligations? It is becoming clearer that like human beings' animals do have certain rights as living beings. All most all religions of the world have prescribed certain rights to them which are encrypted in their holy books. Most of the people are aware of basic concept of observing mild and kind behavior towards pet animals only. Today we scientifically know that humanity and animals have co-existing inevitability. By observing mutual behavior and sensation, the world has provided special rights to the animals by making laws and Acts. Implementation of those rights can only be made through mutual understanding of the citizens under the supervision of law enforcing agencies. It is the responsibility of state and citizens of the states jointly to protect the animals when their rights are being violated.

Enterotoxigenic methicillin resistant Staphylococcus aureus contamination in salted fish from Gwadar Balochistan.

Staphylococcus aureus is an important foodborne pathogen associated to food intoxication and other multiple infections in human being. Its presence in salted food is a serious issue due to its salt tolerance potential. A study was conducted to analyze the presence of enterotoxins producing drug resistance S. aureus in salted sea fish from Gwadar. Freshly persevered samples (n=50) of salted fish were subjected to analyze the presence of S. aureus using 16S rRNA and Nuc genes primers. The isolates were then evaluated for drug resistance and enterotoxins producing potential using specific primers for MecA (methicillin resistance gene), (SEA) staphylococcal enterotoxin A and (SEB) staphylococcal enterotoxin B genes. Total 13/50 (26%) of the samples were found positive for the presence of S. aureus, preliminary confirmed with biochemical profiling and finally with the help of target genes presence. The isolates were found showing 100% resistant to methicillin, which were molecularly confirmed by the presence of MecA gene present in genome. The isolates 5/13 (38%) were positive for SEA and 3/13 (23%) for SEB genes, whereas 2/13 (15%) were confirmed having both SEA and SEB genes in its genome. It was also confirmed that all the isolates were capable to form biofilm over the glass surfaces. It was concluded that the study confirmed the presence of enterotoxigenic methicillin resistance Staphylococcus aurous (MRSA) in salted fish product, that poses gross food safety concern. Preventive and control measures are necessary to handle this serious food safety concern.

Accreditation and evaluation of veterinary medical institutes - a Pakistani perspective.

The Higher Education Commission of Pakistan (HEC) is an independent, autonomous and constitutionally established institution that provides primary funding for higher education in Pakistan, which it also oversees, regulates and accredits. According to the HEC Charter: 'the HEC of Pakistan may set up national or regional evaluation councils or authorise any existing council/or similar body to carry out accreditation of institutions including their departments, facilities and disciplines by giving them appropriate ratings'. At present, 14 accreditation bodies operate under the umbrella of the HEC. Five were established under the HEC's Quality Assurance Agency, while nine councils associated with the accreditation and evaluation of medical institutes are independent bodies that were extant before the HEC was created, and are recognised by the HEC. The authors will discuss the role of the Pakistan Veterinary Medical Council (PVMC), the accreditation of veterinary institutes and associated issues of concern. They suggest that the PVMC should adopt an unbiased approach to new and established veterinary institutes to attain uniform accreditation of all veterinary institutes throughout the country. In addition, the government should consult the accreditation councils and professional bodies before setting up new institutes. The authors also suggest that, instead of placing new veterinary institutes under the aegis of non-veterinary universities, they should be made sub-campuses of established veterinary universities. In time, they will emerge as established institutes in their own right.

La Commission de l'enseignement supérieur du Pakistan (HEC) est une entité indépendante et autonome ayant pour missions constitutionnelles de réunir l'essentiel du financement destiné à l'enseignement supérieur au Pakistan et d'accréditer, superviser et réglementer les institutions d'enseignement supérieur du pays. En vertu de sa Charte, « la HEC du Pakistan peut instituer des Conseils nationaux ou régionaux d'évaluation ou agréer les Conseils ou organismes déjà en place afin de procéder à ces évaluations, dans le but d'accréditer les institutions d'enseignement supérieur y compris au niveau de leurs départements, centres de formation et disciplines, à travers un système de notation approprié ¼. À l'heure actuelle, 14 organismes d'accréditation travaillent sous les auspices de la HEC. Cinq d'entre eux opèrent au sein de l'Agence d'assurance qualité de la HEC tandis que l'accréditation et l'évaluation des établissements d'enseignement du secteur médical sont assurées par neufs Conseils indépendants dont la création est antérieure à celle de la HEC mais que cette dernière a agréés. Les auteurs examinent le rôle joué par le Conseil de la médecine vétérinaire du Pakistan (PVMC), le processus d'accréditation des écoles vétérinaires et les problèmes rencontrés dans ce domaine. Ils préconisent que le PVMC fasse preuve d'impartialité à l'égard des établissements d'enseignement de la médecine vétérinaire, nouveaux ou anciens, afin de garantir l'homogénéité de l'accréditation à l'échelle nationale. Ils recommandent en outre que le gouvernement consulte les Conseils d'accréditation et les ordres professionnels avant toute création de nouvel établissement. Enfin, les auteurs proposent que les nouveaux établissements de formation vétérinaire mis en place ne soient pas placés sous l'égide d'universités généralistes mais enregistrés comme départements nouveaux au sein d'écoles vétérinaires déjà établies. Au fil du temps, ces départements seront reconnus en tant que facultés vétérinaires autonomes.

La Comisión de Educación Superior (HEC) del Pakistán, órgano independiente y autónomo establecido por vía constitucional, es la principal fuente de financiación de la enseñanza superior del país, además de supervisar, regular y acreditar a los establecimientos que dispensan estos estudios. Según reza su Estatuto, la Comisión de Educación Superior del Pakistán puede crear consejos nacionales o regionales de evaluación o habilitar a todo consejo u órgano similar existente para que asuma las labores de acreditación de los establecimiento en cuestión (lo que incluye sus departamentos e instalaciones y las disciplinas impartidas) y los clasifique según corresponda. En la actualidad hay 14 entidades de acreditación que operan bajo la égida de la HEC. Cinco de ellas fueron establecidas desde el Organismo de Garantía de Calidad de la HEC, mientras que otros nueve consejos vinculados a la acreditación y evaluación de institutos de enseñanza médica y veterinaria son entidades independientes que ya existían antes de la creación de la HEC y que esta ha reconocido. Tras exponer la función del Consejo de Medicina Veterinaria del Pakistán (PVMC), los autores presentan el proceso de acreditación de los institutos de enseñanza veterinaria y describen los problemas que se plantean en este terreno. Desde su punto de vista, el PVMC debe aplicar criterios imparciales a los institutos de enseñanza veterinaria, nuevos o ya asentados, para hacer posible una acreditación homogénea de todos estos establecimientos en el conjunto del país. Además, el Gobierno debe consultar a los consejos de acreditación y órganos profesionales antes de establecer nuevos institutos. En opinión de los autores, en lugar de colocar los nuevos institutos de veterinaria bajo la égida de facultades generalistas ajenas a la veterinaria, es preferible constituirlos como subcampus de las facultades de veterinaria existentes, subcampus que con el tiempo se asentarán y adquirirán por derecho propio la condición de institutos de enseñanza veterinaria.

Nasal immunization with inhibin DNA vaccine delivered by attenuated Salmonella choleraesuis for improving ovarian responses and fertility in cross-bred buffaloes.

This study was conducted to determine the effect of immunization with inhibin DNA vaccine delivered by attenuated Salmonella choleraesuis on ovarian responses and fertility in cross-bred buffaloes. A total of 134 cross-bred buffaloes were divided into four groups: groups T1 (n = 34), T2 (n = 35) and T3 (n = 31) were nasal immunized twice a day with 10 ml of 1 × 1010  CFU/ml of the C501 (pVAX-asd-IS) vaccine for 5, 3 and 1 day, respectively. Group C (n = 34) was nasal immunized with 10 ml PBS for 5 days. All animals were immunized twice with an interval of 14 days and administered with 200 µg of a GnRH analogue on day 28, 0.5 mg PGF2α on day 35 and 200 µg of the same GnRH analogue on day 37. TAI was performed at 18 and 24 hr after the second GnRH treatment. Fourteen days after primary immunization, C501 (pVAX-asd-IS) elicited significant immune responses, and anti-inhibin IgG antibody titres in group T1 were significantly higher (p < .01) than groups T3 and C. After the second GnRH treatment, the growth speed of the dominant follicles in group T1 was significantly faster (p < .05) than groups T3 and C. The number and diameter of large follicles (≥10 mm) as well as ovulatory follicles in group T1 were the greatest in all groups, resulting in a greater conception rate in buffaloes with positive anti-inhibin antibodies. These results demonstrate that immunization with the C501 (pVAX-asd-IS) vaccine, coupled with the Ovsynch protocol, could be used as an alternative approach to improve reproductive performance in cross-bred buffaloes.

Excretory/secretory products of adult Haemonchus contortus and Teladorsagia circumcincta which increase the permeability of Caco-2 cell monolayers are neutralised by antibodies from immune hosts.

The onset of abomasal pathophysiology due to parasitism coincides with the presence of adult worms in the lumen, implicating worm excretory/secretory (ES) products acting on the surface mucosa. Caco-2 cell monolayers were grown to confluence on Transwell plates and exposed on the apical side to ES products of adult Haemonchus contortus and Teladorsagia circumcincta. ES products of both species significantly (p<0.001) reduced transepithelial electrical resistance after 2h to 81.1±1.0% and 82.9±1.1% respectively. Immunocytochemical staining of the Caco-2 monolayers for zona occludens-1 and occludin confirmed that the tight junctions remained intact in control medium, but these proteins were internalised from disrupted junctions after exposure to ES products. The components of H. contortus ES products responsible for increased epithelial permeability were partially blocked by phage displaying single chain antibodies derived from sheep immune to field infection and enriched by panning with H. contortus ES products. Immune hosts may therefore be able to reduce the effects of worm chemicals on the gastric epithelium. Permeabilisation of the abomasal surface mucosa by worm chemicals would also explain how cells deep in the gastric glands could rapidly be affected by parasites emerging from the glands or within a day of transplantation of adult worms into naïve hosts, resulting in the pathophysiology typically caused by abomasal nematode parasitism.

Genetics of human Bardet-Biedl syndrome, an updates.

Bardet-Biedl syndrome (BBS) is an autosomal recessive multisystemic human genetic disorder characterized by six major defects including obesity, mental retardation, renal anomalies, polydactyly, retinal degeneration and hypogenitalism. In several cases of BBS, few other features such as metabolic defects, cardiovascular anomalies, speech deficits, hearing loss, hypertension, hepatic defects and high incidence of diabetes mellitus have been reported as well. The BBS displays extensive genetic heterogeneity. To date, 19 genes have been mapped on different chromosomes causing BBS phenotypes having varied mutational load of each BBS gene. In this review, we have discussed clinical spectrum and genetics of BBS. This report presents a concise overview of the current knowledge on clinical data and its molecular genetics progress upto date.

Immunomodulatory effects of adult Haemonchus contortus excretory/secretory products on human monocyte-derived dendritic cells.

The levels of expression of surface molecules and release of cytokines and chemokines of human monocyte-derived dendritic cells were determined after their exposure to adult H. contortus excretory/secretory (ES) products or a combination of ES products and bacterial lipopolysaccharide (LPS). Worm products provoked a weak response and only partial maturation of the dendritic cells, consistent with the hyporesponsiveness and more tolerogenic immune environment present in parasitized animals and humans. Co-stimulation with LPS demonstrated that H. contortus secretions, like those of other helminths, contain immunomodulators capable of reducing some aspects of the strong T(H)1/T(H)2 response evoked by bacterial LPS. There were significant reductions in the release of some cytokine/chemokines by LPS-stimulated mdDCs and a trend (although not significant at P < 0.05) for reduced expression levels of CD40, CD80 and HLA-DR. A prominent feature was the variability in responses of dendritic cells from the four donors, even on different days in repeat experiments, suggesting that generalized conclusions may be difficult to make, except in genetically related animals. Such observations may therefore be applicable only to restricted populations. In addition, previous exposure to parasites in a target population for immunomodulatory therapy may be an important factor in assessing the likelihood of adverse reactions or failures in the treatment to worm therapy.

Value of D-dimers assay in diagnosis of pulmonary embolism.

OBJECTIVE: To determine the value of D-Dimers assay in the diagnosis of Pulmonary Embolism (PE) at Armed Forces Institute of Pathology, Rawalpindi, Pakistan from January to November 2000. METHODS: Forty three consecutive patients clinically suspected of suffering from pulmonary embolism and referred to Armed Forces Institute of Pathology for Ventilation-Perfusion scan of lungs were inducted in the study. A detailed history was taken and clinical examination was performed. All patients were then subjected to Perfusion and/or Ventilation scan, which was taken as the standard for the diagnosis of PE. Blood samples were collected from all patients in trisodium citrate. Platelet poor plasma obtained from the samples was tested for D-Dimers semi-quantitatively using latex agglutination method. RESULTS: Out of 43 patients 14 (32.6%) had completely normal ventilation-perfusion scan hence the diagnosis of PE was excluded. In 6 (13.9%) patients the results were considered indeterminate. Abnormalities suggestive of pulmonary embolism were detected in 23 (53.5%) patients. D-Dimers were less than the cut off value of 500 ng/ml in 19 (44.2%) patients, whereas in 24 (55.8%) the levels were more than 500 ng/ml. When compared with the results of ventilation-perfusion scan the sensitivity of D-Dimers was 91.3% and specificity was 100%. Positive predictive value of the test was 100% whereas negative predictive value was 87.5%. CONCLUSION: D-Dimers assay combined with high clinical evidence is a cost effective, readily available test which can safely exclude the diagnosis of pulmonary embolism in majority of the cases.

Biochemical changes in wheat during storage at three temperatures.

Biochemical changes in wheat grains stored at 10, 25 and 45 degrees C for six months were studied. A significant decrease in pH and an increase in titratable acidity was observed during storage of wheat grains at 25 degrees C and 45 degrees C. Moisture contents of wheat grains decreased by 15% at 25 degrees C and 26% at 45 degrees C during six months of storage. A significant decrease in water soluble amylose (20-28%) along with an increase in insoluble amylose contents (7.6-17%) were observed during storage at 25 and 45 degrees C. Amylase activity of the samples showed a decrease as the storage progressed. Total soluble sugars increased by 9% at 10 degrees C and 12% at 25 degrees C; a 37% decrease was observed after six months storage at 45 degrees C. Total available lysine decreased by 18.0% and 22.6% at 25 and 45 degrees C, respectively, after six months storage. In vitro protein digestibility of wheat grains decreased by 5.00% at 25 degrees C and 10.28% at 45 degrees C during six months of storage. However, no significant biochemical changes occurred during storage at 10 degrees C.