Hepatic Responses of A Freshwater Fish Against Aquatic Pollution*

Abstract.-Freshwater fish, Tor putitora caught from polluted portion of River Kabul was studied for various hepatic biochemical parameters and was compared with control fish caught from non polluted Warsak Dam to know the possible toxic effects of pollution in the river. Fish liver was dissected out and analyzed for various biochemical parameters like total protein, soluble protein, total cholesterol, total lipid, glucose, free amino acid, DNA, RNA and enzymes such as amylase, GOT, GPT and LDH. The hepatic total proteins, soluble proteins, total cholesterol, total lipids, glucose content and free amino acids in fish sample 1 and sample 2 increased 29% and 16%, 6% and 15%, 75% and 68%, 41% and 65%, 47% and 26%, and 333% and 729%, respectively as compared to control sample. The DNA content decreased 14% and 20%, whereas the RNA decreased 41% and 32% in sample 1 and 2, respectively. The fish sample from site I and II showed 5% and 17% increased hepatic amylase activity, 69% and 72% decreased GOT activity in sample 1 and 2, respectively. The GPT activity decreased 45% in sample 1 and increased 28% in sample 2. The LDH activity was decreased 69% in sample 1, but showed 14% increase in sample 2 as compared to control sample. The increase and decrease in various biochemical parameters and enzymes in the liver of test fish samples in comparison with the control shows the adverse effect of aquatic pollution on the fish health. Aquatic pollution could be one of the major reasons of rapid decline in population of this endangered South Asian fresh water fish

Fish white muscle as biomarker for riverine pollution.

Abstract.-An endangered South Asian freshwater fish, Tor putitora netted from polluted part of River Kabul was studied for various muscle biochemical parameters and was compared with control fish caught from non polluted Warsak Dam water reservoir to investigate the remedies caused by the ambient pollution in the fish health. Fish muscle was analyzed for various biochemical parameters like total protein, soluble protein, total cholesterol, total lipid, glucose, free amino acid, DNA, RNA and free fatty acids and enzymes such as amylase, GOT, GPT and LDH. The muscles showed an increase in total muscle proteins (11.8% and 89.6%), total lipid (406.8% and 119.6%), DNA (18.7% and 18.8%) in sample 1 and 2 respectively. Soluble protein, however, remained unaffected in sample 1 but showed a drastic 77.35% increase in sample 2. The muscles showed a decrease in total cholesterol (17.22% and 12.43% ), glucose (41.26% and 51.00%) and RNA (23.95% and 40.04%) in sample 1 and sample 2 respectively. However, free amino acids decreased 12.14% in sample 1 and increased 31.3% in sample 2. Similarly free fatty acids decreased 53.5% in sample 1 but showed 97% higher value in sample 2. Among enzymes amylase (36.3% and 42.8%), LDH (21.8% and 45.8% ) increased, likewise GOT activity ( 18.72% and 43.34%) and GPT (45.82% and 48.6%) decreased in sample 1 and 2, respectively, as compared with that of control fish. The increase and decrease in various biochemical parameters and enzymes in the white muscle of test fish samples in comparison with the control, reveals the adverse effect of aquatic pollution on the inhabitant fish health. Pollution stress could be one of the reasons of rapid decline in population of this South Asian endangered freshwater fish

Bioaccumulation of hexavalent chromium in commercially edible fish grass carp, Ctenopharyngodon idella

The current research was conducted with the aim to investigate the rate of chromium accumulation in various tissues of commercially valuable freshwater fish, Grass Carp, Ctenopharyngodon idella. Investigation included various tissues of skin, muscles, liver, swim bladder, intestine, and gills in fish. For this purpose, the fish were exposed to sub lethal concentration of hexavalent chromium in the form of potassium dichromate with a dose of 120 mg/L for 40, 20 days and 10 mg/L for 7, 25, 40 and 60 days. Forty fish of the same weight and length (70.45±2.91 g and 7.32±0.16 inches) were used in the present research. Chromium was estimated after acid digestion of the sample tissues and further analyzed through atomic absorption spectrophotometer (Spectra AA6300 China). The results showed that in all the experiments intestine remained the highly accumulative tissue and the accumulation of chromium in the intestine increased with the increase in exposure time i.e., 0.63±0.21 after 7 days, 0.83±0.35 after 25 and 1.63±0.44 after 60 days. Finding of this research work supports that the route of metal uptake was mainly oral instead of absorption by gills or skin

Fresh Records on Water Quality and Ichthyodiversity of River Swat at Charsadda, Khyber Pakhtunkhwa

Abstract.-A study on the fish fauna and water quality parameters of River Swat at Charsadda, Khyber Pakhtunkhwa, Pakistan was conducted, with an aim to determine fresh records of water quality variables and fish fauna after heavy floods of July, 2010 in the province. For fish distribution a 35 km belt of the river was explored for four months and every effort was made to collect the fish specimen by any legal mean. During this study the mean values recorded for physico-chemical parameters were water temperature 19.8°C, pH 7.6, dissolved oxygen 9.65 mg/l, electrical conductivity 199.47 µS/cm, TDS 127.66 mg/l, TSS 93.78 mg/l, total hardness 118.75 mg/l, calcium hardness 75 mg/l, magnesium hardness 43.75 mg/l, sodium 5.325 mg/l, potassium 3.175 mg/l, total alkalinity 97 mg/l, chloride 15.3 mg/l and nitrite 0.0092 mg/l. All these values were within the limits prescribed by the standard methods for the examination of water, sewage and industrial wastes. In the present study a total of 38 fish species belonging to; 6 orders, 9 families and 24 genera were recorded. Cyprinidae was the richest family and was represented by 20 species, Nemacheilidae by 4, Sisoridae by 6, Channidae and Schilbidae by 2, Mastacembelidae, Schilbidae, Belonidae and Chandidae by single species. Results report six (6) species missing in the present study in comparison with the past records

Poliovirus excretion among children with primary immune deficiency in Pakistan: A pilot surveillance study protocol

Introduction: Children with primary immunodeficiency disorders (PID) are more susceptible to developing viral infections and are at a substantially increased risk of developing paralytic poliomyelitis. Such children, if given oral polio vaccines tend to excrete poliovirus chronically that may lead to the propagation of highly divergent vaccine-derived poliovirus (VDPV). Consequently, they may act as a reservoir for the community by introducing an altered virus potentially imposing a risk to global polio eradication. However, the risks of chronic and prolonged excretion are not well characterised in the study context. This study seeks to establish a pilot surveillance system for successful identification and monitoring of VDPV excretion among children with PID. It will assess whether the Jeffrey Modell warning signs of PID can be used as an appropriate screening tool for PID in Pakistan.Methods and analysis: In this pilot surveillance, recruitment of PID cases is currently done at participating hospitals in Pakistan. Potential children are screened and tested against the Jeffrey Modell Foundation (JMF) warning signs for immunodeficiency and their stool is collected to test for poliovirus excretion. Cases excreting poliovirus are followed until the two consecutive negative stool samples are obtained over a period of 6 months. The data will be analysed to calculate hospital-based proportions of total Immunodeficiency-related vaccine-derived poliovirus (iVDPV) cases over a 2-year period and to determine the sensitivity and specificity of the JMF signs.Ethics and dissemination: This protocol was reviewed and approved by the WHO (WHO Reference-2018/811124-0), Aga Khan University (AKU ERC-2018-0380-1029) and National Bioethics Committee (Ref No. 4-87 NBC-308-Y2). The results will be published in an open access peer-reviewed scientific journal and presented to the iVDPV Working Group members, policy-makers, paediatric consultants and fellow researchers with the same domain interest. It may be presented in scientific conferences and seminars in the form of oral or poster presentations

Evaluation of vaccine derived poliovirus type 2 outbreak response options: A randomized controlled trial, Karachi, Pakistan

Background: Outbreaks of circulating vaccine derived polioviruses type 2 (cVDPV2) remain a risk to poliovirus eradication in an era without live poliovirus vaccine containing type 2 in routine immunization. We evaluated existing outbreak response strategies recommended by the World Health Organization (WHO) for control of cVDPV2 outbreaks.Methods: Seronegative children for poliovirus type 2 (PV2) at 22 weeks of life were assigned to one of four study groups and received respectively (1) one dose of trivalent oral poliovirus vaccine (tOPV); (2) monovalent OPV 2 (mOPV2); (3) tOPV together with a dose of inactivated poliovirus vaccine (IPV); or (4) mOPV2 with monovalent high-potency IPV type 2. Stool and blood samples were collected and assessed for presence of PV2 (stool) and anti-polio antibodies (sera).Results: We analyzed data from 265 children seronegative for PV2. Seroconversion to PV2 was achieved in 48, 76, 98 and 100% in Groups 1–4 respectively. mOPV2 was more immunogenic than tOPV alone (p \u3c 0.001); and OPV in combination with IPV was more immunogenic than OPV alone (p \u3c 0.001). There were 33%, 67%, 20% and 43% PV2 excretors in Groups 1–4 respectively. mOPV2 resulted in more prevalent shedding of PV2 than when tOPV was used (p \u3c 0.001); and tOPV together with IPV resulted in lower excretion of PV2 than tOPV alone (p = 0.046).Conclusion: mOPV2 was a more potent vaccine than tOPV. Adding IPV to OPV improved immunological response; adding IPV also seemed to have shortened the duration of PV2 shedding. mIPV2 did not provide measurable improvement of immune response when compared to conventional IPV. WHO recommendation to use mOPV2 as a vaccine of first choice in cVDPV2 outbreak response was supported by our findings

Effectiveness of typhoid conjugate vaccine against culture-confirmed salmonella enterica serotype typhi in an extensively drug-resistant outbreak setting of Hyderabad, Pakistan: A cohort study

Background: Salmonella enterica serotype Typhi (S Typhi) is a major public health problem in low-income and middle-income countries. We aimed to investigate the effectiveness and impact of the typhoid conjugate vaccine Typbar-TCV against S Typhi among children in an outbreak setting of extensively drug-resistant (XDR) S Typhi in Pakistan.Methods: This cohort study was done from Feb 21, 2018, to Dec 31, 2019. A census survey of all households located in the Qasimabad and Latifabad subdistricts of Hyderabad, Pakistan, was done at baseline, and 174 005 households were registered in the census. The Typbar-TCV immunisation campaign was initiated at temporary vaccination centres and 207 000 children aged 6 months to 10 years were vaccinated from Feb 21, 2018, to Dec 31, 2018. Social mobilisers informed parents about the vaccination process. Vaccination records were maintained electronically and linked with the household census surveys. Active surveillance for suspected and blood-culture-confirmed S Typhi was established in hospitals, clinics, and laboratories to assess the following outcomes: cases of suspected typhoid fever, culture-confirmed S Typhi, and antimicrobial resistance. An age-stratified cohort of 1100 vaccinated children was randomly selected from the vaccination registry, tested for Vi-IgG antibodies (data not reported), and followed up fortnightly (via telephone calls or household visits) until Dec 31, 2019, for ascertainment of outcomes during the study period. 20 847 vaccinated and unvaccinated children were randomly selected from the census registry as a quality control cohort and followed up from Oct 1 to Dec 31, 2019, for ascertainment of outcomes. Vaccine effectiveness against suspected, culture-confirmed, and XDR S Typhi was calculated.Findings: 23 407 children from the census registry and surveillance system were included in the vaccine effectiveness analysis. 13 436 (57·4%) children were vaccinated, 12 214 (52·2%) were male, and 10 168 (43·4%) were aged 6-59 months. 5378 (23·0%) of 23 407 children had suspected S Typhi, among whom 775 (14·4%) had culture-confirmed S Typhi and 361 (68·6%) of 526 had XDR S Typhi. Vaccine effectiveness was 55% (95% CI 52-57) against suspected S Typhi (regardless of culture confirmation), 95% (93-96) against culture-confirmed S Typhi, and 97% (95-98) against XDR S Typhi.Interpretation: Typbar-TCV is effective in protecting children against S Typhi infection in an outbreak setting, and was able, with moderate deployment, to curtail a major XDR S Typhi outbreak in a densely populated setting. The vaccine shows efficacy against S Typhi irrespective of antimicrobial resistance.Funding: Bill & Melinda Gates Foundation

Effectiveness of 10-valent pneumococcal conjugate vaccine against vaccine-type invasive pneumococcal disease in Pakistan

Objective: To assess the effectiveness of 10-valent pneumococcal conjugate vaccine (PCV10) against invasive pneumococcal disease (IPD) due to vaccine serotypes of Streptococcus pneumoniae post introduction of the vaccine into the routine immunization program in Pakistan.Methods: A matched case-control study was conducted at 16 hospitals in Sindh Province, Pakistan. Children aged (eligible to receive PCV10) who presented with radiographically confirmed pneumonia and/or meningitis were enrolled as cases. PCR for the lytA gene was conducted on blood (for radiographic pneumonia) and cerebrospinal fluid (for meningitis) samples to detect S. pneumoniae. The proportion of IPD due to vaccine serotypes (including vaccine-related serogroups) was determined through serial multiplex PCR. For each case, at least five controls were enrolled from children hospitalized at the same institution, matched for age, district, and season.Results: Of 92 IPD patients enrolled during July 2013 to March 2017, 24 (26.0%) had disease caused by vaccine serotypes. Most case (87.5% of 24) and control (66.4% of 134) children had not received any PCV10 doses. The estimated effectiveness of PCV10 against vaccine-type IPD was 72.7% (95% confidence interval (CI) -7.2% to 92.6%) with at least one dose, 78.8% (95% CI -11.9% to 96.0%) for at least two doses, and 81.9% (95% CI -55.7% to 97.9%) for all three doses of vaccine.Conclusion: The vaccine effectiveness point estimates for PCV10 were high and increased with increasing number of doses. However, vaccine effectiveness estimates did not reach statistical significance, possibly due to low power. The findings indicate the likely impact of vaccine in reducing the burden of vaccine-type IPD if vaccine uptake can be improved

Determinants of response at 2 months of treatment in a cohort of Pakistani patients with pulmonary tuberculosis

Mycobacterium tuberculosis infection continues to be a major global challenge. All patients with pulmonary tuberculosis are treated with a standard 6-month treatment regimen. Historical data suggest that even with shortened treatment, most patients achieve long-term remission. Risk stratification is a goal for reducing potentially toxic prolonged treatment. This study aimed to determine the factors associated with the early clearance of sputum acid-fast bacilli (AFB). A total of 297 freshly diagnosed patients with pulmonary tuberculosis were included and enrolled in this study. Information related to their ethno-demographic and anthropometric characteristics was collected. We also assessed their complete blood counts, and blood iron, folate, and vitamin B12 levels. We found that the presence of higher levels of acid-fast bacilli (AFB) in diagnostic sputum microscopy was the single most significant prognostic factor associated with early clearance of sputum AFB after 2 months of treatment. All of our patients achieved treatment success after 6 months of treatment and were disease free. Our results support the data obtained from previous studies indicating that AFB clearance at 2 months is unlikely to be a clinically useful biomarker or indicator for therapeutic stratification. Furthermore, demographic, anthropometric, and nutritional factors are not clinically useful biomarkers

Pollution of Large, Subtropical Rivers-River Kabul, Khyber-Pakhtun Khwa Province, Pakistan) : Physico-Chemical Indicators

Abstract.-Water quality in River Kabul was monitored to determine if indusatrial effluents and city sewage had a significant impact on it. Water samples were collected three times a year, two times in winter (low Q) and once during summer (high Q) for three years. Sampling was carried out along a section of River Kabul receiving untreated industrial effluents to assess the physico-chemical variables and was compared with upstream water from Warsak Dam being pristine. Factories discharge effluents having high organic pollutants load into the river which is evident from the high value (290.5, 280 mgO 2 /l for low Q and 295.7, 281.7 mgO 2 /l for high Q periods) of biological oxygen demand (BOD) in comparison with the National Environmental Quality Standard (NEQS) value of 80 mgO 2 /l set for this variable. The river water downstream of entry of industrial wastewater had total suspended solids, biological oxygen demand, NH 3 , Cl -and SO 4 2-17%, 8%, 58%, 42%, 17% and 148% higher than those in the upstream water during low Q (winter), whereas these variables were 6%, 100%, 286%, 25% and 132% higher than those in upstream water, respectively, during the high Q (summer) season. These raises concern over the long-term health of the River Kabul ecosystem