Gut microbial ecology and exposome of a healthy Pakistani cohort.

BACKGROUND: Pakistan is a multi-ethnic society where there is a disparity between dietary habits, genetic composition, and environmental exposures. The microbial ecology of healthy Pakistani gut in the context of anthropometric, sociodemographic, and dietary patterns holds interest by virtue of it being one of the most populous countries, and also being a Lower Middle Income Country (LMIC). METHODS: 16S rRNA profiling of healthy gut microbiome of normo-weight healthy Pakistani individuals from different regions of residence is performed with additional meta-data collected through filled questionnaires. The current health status is then linked to dietary patterns through [Formula: see text] test of independence and Generalized Linear Latent Variable Model (GLLVM) where distribution of individual microbes is regressed against all recorded sources of variability. To identify the core microbiome signature, a dynamic approach is used that considers into account species occupancy as well as consistency across assumed grouping of samples including organization by gender and province of residence. Fitting neutral modeling then revealed core microbiome that is selected by the environment. RESULTS: A strong determinant of disparity is by province of residence. It is also established that the male microbiome is better adapted to the local niche than the female microbiome, and that there is microbial taxonomic and functional diversity in different ethnicities, dietary patterns and lifestyle habits. Some microbial genera, such as, Megamonas, Porphyromonas, Haemophilus, Klebsiella and Finegoldia showed significant associations with consumption of pickle, fresh fruits, rice, and cheese. Our analyses suggest current health status being associated with the diet, sleeping patterns, employment status, and the medical history. CONCLUSIONS: This study provides a snapshot of the healthy core Pakistani gut microbiome by focusing on the most populous provinces and ethnic groups residing in predominantly urban areas. The study serves a reference dataset for exploring variations in disease status and designing personalized dietary and lifestyle interventions to promote gut health, particularly in LMICs settings.

In Silico and In Vitro Analysis of Helicobacter pullorum Type Six Secretory Protein Hcp and Its Role in Bacterial Invasion and Pathogenesis.

Helicobacter pullorum is a human zoonotic pathogen transmitted through poultry where it is associated with vibrionic hepatitis and colitis. Hemolysin co-regulated protein (Hcp) is an important structural as well as effector protein of type six secretory system; however, its role in H. pullorum invasion and pathogenesis has not been elucidated. In this study, we predicted the Helicobacter pullorum Hcp (HpuHcp) structure and identified Campylobacter jejuni Hcp (CjHcp) as its nearest homologue. Analysis of the predicted structure shows several common bacterial Hcp motifs like Protein kinase C phosphorylation site, Casein kinase II phosphorylation site, N-myristoylation site, cAMP-and cCGMP-dependent protein kinase phosphorylation site, N-glycosylation site. The presence of unique microbodies C-terminal targeting signal domain was present in HpuHcp which was seen for the first time in CjHcp. This could indicate that Hcp is a structural protein as well as a secretory protein. Moreover, the presence of a deamidase domain, similar to the tecA of Burkholderia cenocepacia an opportunistic pathogen, may help in bacterial internalization as it depolymerises the membranous actin by deamidation of the host cell Rho GTPases cdc42 and Rac1, which was supported by increased invasion of hepatocytes by Hcp-positive isolates.

Morpho-palynological assessment of some species of family Asteraceae and Lamiaceae of District Bannu, Pakistan on the bases of light microscope & scanning electron microscopy.

Pollen micro-morphological features have proven to be helpful for the plant taxonomists in the identification and classification of plants. The aim of this study was to evaluate the palynological features of family Asteraceae and Lamiaceae from flora of District Bannu, Khyber Pakhtunkhwa, Pakistan using both scanning electron microscopy (SEM) and light microscope (LM) for their taxonomic importance. Pollen of seven Asteraceous species belonging to four genera and four Lamiaceae species categorized into four genera were collected from different localities of research area. The present research work provides detailed information of diverse morpho-palynological characters both qualitatively and quantitatively including pollen shape, type, diameter, P/E ratio, exine sculpturing and thickness. Type of pollen in Asteraceae and Lamiaceae was ranged from tricolporate, tricolpate, trizonocolpate and hexazonocolpate. The maximum polar diameter (40.05 µm) and equatorial diameter (37.66 µm) was observed in the Ajuga bracteoosa while minimum polar and equatorial diameter was noted in Isodon rugosus (11.10 µm) and Erigeron canadensis (13.20 µm) respectively. Sculpturing of exine include; echinate, reticulate scabrate, aerolate, reticulate-verrucate, reticulate-scabrate, perforate and reticulate to perforate. Exine thickness was examined maximum 1.50 µm in Helianthus tuberosus, whereas minimum in Conyza Canadensis (0.16 µm). The pollen fertility was found highest in C Canadensis (83.33%) and lowest in Ajuga bracteosa (58.06%). The observed pollen morphology has many valuable qualitative and quantitative attributes for the better understanding of their taxonomy and play significant role in correct identification.

Characterization of the Gastrointestinal and Reproductive Tract Microbiota in Fertile and Infertile Pakistani Couples.

The human microbiota is recognized as a vital "virtual" organ of the human body that influences human health, metabolism, and physiology. While the microbiomes of the gut, oral cavity, and skin have been extensively studied in the literature, relatively little work has been done on characterizing the microbiota of the human reproductive tract organs, and specifically on investigating its association to fertility. Here, we implemented a 16S ribosomal RNA (rRNA) amplicon sequencing approach to sequence and characterize the gut and genital tract microbiomes from several married Pakistani couples. The recruited individuals included 31 fertile and 35 infertile individuals, with ages ranging from 19-45 years. We identified several fluctuations in the diversity and composition of the gut and genital microbiota among fertile and infertile samples. For example, measures of α-diversity varied significantly between the genital samples donated by fertile and infertile men and there was overall greater between-sample variability in genital samples regardless of gender. In terms of taxonomic composition, Actinobacteria, Bacteroidetes, and Firmicutes fluctuated significantly between the gut microbiomes of fertile and infertile samples. Finally, biomarker analyses identified features (genera and molecular functions and pathways) that differed significantly between the fertile and infertile samples and in the past have been associated with bacterial vaginosis. However, we emphasize that 16S amplicon data alone has no bearing on individual health and is merely representative of microbial taxonomic differences that could also arise due to multiple other factors. Our findings, however, represent the first effort to characterize the microbiome associated with fertile and infertile couples in Pakistan and will hopefully pave the way for more comprehensive and broad-scale investigations in the future.

Initial Sequencing and Characterization of the Gastrointestinal and Oral Microbiota in Urban Pakistani Adults.

We report the initial characterization of the gastrointestinal tract (gut) and oral microbiota (bacteria) in 32 urban Pakistani adults. Study participants were between ages 18 and 40, had body mass index between 18 and 25 Kg/m2, and were students or early-career professionals. These individuals donated a total of 61 samples (32 gut and 29 oral) that were subjected to 16S ribosomal RNA (rRNA) gene sequencing. Microbiome composition of Pakistani individuals was compared against the uBiome database of selected individuals who self-reported to be in excellent health. We observed strong gender-based differences in the gut microbiome of Pakistani individuals, a skewness toward Firmicutes, and unusually high levels of Proteobacteria in the Pakistani men. These observations may indicate microbiota dysbiosis, though 16S data alone can neither establish cause nor effect to human health. Albeit conducted on a smaller scale, our report provides a first snapshot about the composition and diversity of gut and oral microbiota communities in Pakistani individuals.

Prevalence and role of Type six secretion system in pathogenesis of emerging zoonotic pathogen Helicobacter pullorum from retail poultry.

Helicobacter pullorum is an emerging zoonotic pathogen that causes gastroenteritis in chickens and inflammatory bowel disease in humans ingesting contaminated meat. However, the mechanism by which the bacterium causes disease is unclear. Type six secretion system (T6SS) plays a major role in bacterial pathogenesis and adaptation. Haemolysin coregulated protein (Hcp) plays a central role in the structure of the T6SS pilus and acts as effector protein in certain bacteria. In this study, H. pullorum isolated from 156 caecal samples of broiler chickens was screened for the presence of T6SS Hcp gene via PCR amplification. 30.7% of caecal and 18.3% of liver samples tested positive for presence of H. pullorum. From these positive samples, 29.7% possessed the T6SS gene. In bacterial co-culture experiments, significant loss of viability (81.6-39.1%) was observed for H. pullorum-infected hepatocytes and presence of Hcp did not contribute to the loss of cell viability. Nevertheless, infection of erythrocytes with Hcp-positive isolates was associated with greater haemolytic activity compared to infection with Hcp-negative isolates. Therefore, presence of T6SS could be indicative of virulent strains meriting further studies to characterize this virulence factor in H. pullorum infection.

Electrolytes imbalance caused by amikacin in patients receiving multi drug resistance- tuberculosis treatment at Hazara region Kpk, Pakistan.

INTRODUCTION: Multi drug resistance-tuberculosis (MDR-TB) is considered amongst one of the fatal medical and public health issue. Greater frequency of adverse reactions has been observed using 2nd line antituberculosis drugs, which rises the rate of morbidity. Hypokalemia and hyponatremia are common in patients receiving MDR-TB treatment. MATERIALS AND METHODS: This study was aimed to assess the effect of amikacin as multidrug resistance tuberculosis treatment on serum sodium and potassium level. A total 179 subjects (82 males and 97 females) were screened within the period of one year for their serum electrolyte levels before and during the treatment. Their body weight and sociodemographic characteristics were also taken into considerations. Data were expressed in the form of mean ± standard error using One-way analysis of variance (ANOVA), followed by Dunnett's test. All the statistical analyses were performed using GraphPad Prism v. 6. RESULT: The results showed that amikacin caused a significant (p< 0.05) hypokalemia when used in combination with other drugs having no known effect on serum potassium level which proved to be the side effect of amikacin. The average serum potassium level in both intensive and continuation phase was significantly lower (p< 0.05) than the baseline (3.73 mmol/L), however, in the continuation phase the potassium level (3.58 mmol/L) was slightly higher than that of intensive phase (3.54 mmol/L). The average serum sodium level in both phases; intensive and continuation, remained in steady state (137.25 mmol/L and 137.87 mmol/L respectively) throughout the study period as compared to the base line (135.5 mmol/L). The amikacin drug lowered serum potassium level but had no effect on sodium level. CONCLUSIONS: It is concluded that the serum electrolytes should be monitored during the course of treatment and corrected accordingly.

Helicobacter pullorum: An Emerging Zoonotic Pathogen.

Helicobacter pullorum (H.pullorum) commonly colonizes the gastrointestinal tract of poultry causing gastroenteritis. The bacterium may be transmitted to humans through contaminated meat where it has been associated with colitis and hepatitis. Despite the high prevalence of H. pullorum observed in poultry, little is known about the mechanisms by which this bacterium establishes infection in host and its virulence determinants. In this article we aim to provide an overview of this emerging zoonotic pathogen; its general characteristics, hosts, prevalence, and transmission as well as its pathogenic potential. We also discuss possible control strategies and risk of disease emergence.

Anti-Infective Potential Of A Medicinal Plant Pimpenella Stewartii Used Traditionally In Hazara.

BACKGROUND: Medicinal plants have been used since long as folk medicines and also in Ayuvedic and Unani systems of treatment. They have importance to save human and animal lives because of their utilization in the herbal medicines. In all over the word plants are in constant use for curing of various kinds of diseases. The herbal medicines are effective due to presence of bioactive compounds along with vitamins and minerals in them. Moreover, plants produce photo-chemicals mainly secondary metabolites which have significant pharmaceutical activities like antioxidant, antimicrobial, pesticidal and anti-tumour activity. They are very effective and have little or no side effects. METHODS: Five microbial strains including three fungal and two bacterial species were tested for the efficacy of plant materials by Agar diffusion method. The fungal species were Aspergilus flavus, Aspergilus niger and Alternaria alternata while bacterial species included Arvenia caratovora and Xanthomans spp. RESULTS: Results obtained for methanolic extracts of Pimpenella stewartii against Aspergilus flavus, Aspergilus niger and Alternaria alternata showed significant antifungal activity. The antibacterial screening of Methanolic extracts of Pimpenella stewartii were evaluated against Arvenia caratovora and Xanthomans spp. The best zone of inhibition was found against Xanthomans spp (97.33±2 mm) at 250 ppm concentration followed by Arvenia caratovora (49.7±14 mm) at same concentration.