A staggering rise of HIV cases in Pakistan

In the last decade,Pakistan has experienced multidemics of HIV in key populations,namely:injecting drug users,male sex with male,female sex workers,transgender sex workers,and transgenders.According to recent reports,in Pakistan,210000 people with HIV accounts for less than 0.2%of the general population.

Phenotypic and Antimicrobial Susceptibility Studies of Proteus mirabilis Isolates from Fresh Water Fishes in FCT, Abuja-Nigeria

Our study was carried out to determine the phenotypic characterization antimicrobial susceptibility of Proteus mirabilis from fish in FCT, Abuja using isolation, selective plating, preliminary observation, complete biochemical method and antimicrobial susceptibility testing. The biochemical tests conducted includes include Citrate Utilization test, Triple Sugar Iron test, Urea test, Methyl Red test, Indole test, and Voges Proskauer test. The isolates were confirmed by MicrobactTMGNB24E identification kit (Oxiod, UK). A total of 400 fish samples were bought in the market from three area council of the FCT. The result of the study showed overall prevalence rate of (13) 3.25% of Proteus mirabilis isolates. Distribution based on Area councils showed that AMAC had higher prevalence rate of 4.81%, while Bwari had 2.99% and Gwagwalada with 2.57% prevalence. All isolates were subjected to antimicrobial susceptibility testing using the modified single disc diffusion method. From the antimicrobial susceptibility testing done it was discovered that Proteus mirabilis are resistant to Amoxyclav (100%), Erythromycin (92.3%), Tetracycline (92.3%) and Ceftriaxone (23.1%). However, the isolates were susceptible to Ofloxacin (100%), Netillin (92.3%), Levofloxacin (92.3%), Ceftazidime (76.9%), Co-trimoxazole (69.2%) and Gentamicin (61.5%). Since Proteus mirabilis sources of zoonotic diseases and can potentially be dangerous to humans and other animals, our research was able to isolate it from fresh water fish sold in the Federal Capital Territory. This makes public health awareness of the risks associated with Proteus mirabilis in Nigeria necessary.

The assessment of good and bad bacteria in holy basil(Ocimum sanctum)leaves

Holy basil(Tulsi,Ocimum sanctum)is considered to be a pious plant.It is used not only in many of the rituals of Hindus but is also known for its innumerable medicinal qualities.Holy basil leaves(HBLs)are often consumed raw thus it is pertinent to detect the bacteria on Tulsi leaves.This study was conducted to understand the diversity of the aerobically growing microbiome of edible holy basil leaves.A total of 112 samples of leaves were collected from households of six localities in and around Bareilly,India for detecting culturable bacteria growing aerobically.A sum of 579 bacterial isolates belonging to 106 different species was identified.Pantoea agglomerans isolates were detected in 43 samples followed by isolates Virgibacillus pantothenticus(21),Bacillus coagulans(18),Bacillus cereus(17),Geobacillus stearothermophilus(16),Klebsiella pneumoniae(13),Citrobacter freundii(12),Lysinibacillus sphaericus(12),Acinetobacter calcoaceticus(11),Escherichia coli(11),and Xenorhabdus bovienii(11).Isolates of 95 species of bacteria were detected on<10 samples of HBLs.Of 579 isolates,161 isolates detected in 68(60.71%)samples were of putatively good bacteria belonging to eight genera(Bacillus,Brevibacillus,Lysinibacillus,Paenibacillus,Photobacter,Siccibacter,Virgibacillus,Xenoirhabdus)and 23 species.However,418 bacterial isolates detected in 108(96.43%)of the HBL samples were classified as potentially pathogenic for humans and animals belonging to 34 genera and 73 species.There were only four samples(three from Mahanagar and one from Bhojipura)that had no potential pathogenic bacteria detected in this study.The analysis indicated that HBL leaves from the ICAR-IVRI campus had a lower probability(P<0.04)of harbouring good bacteria than samples from Bhojpura,Mahanagar,North City and Rajendra Nagar.Cinnamaldehyde inhibiting 96.72%of the isolates and carvacrol inhibiting 96.37%of the isolates were the best herbal antimicrobials and among antibiotics,meropenem inhibited 98.96%of bacterial isolates.Antimicrobial resistance in non-pathogenic or good bacteria was significantly more common(P<0.01)for ajowan oil but they were more often(P<0.05)susceptible to holy basil oil,lemongrass oil,citral,P.embalica leaf extract,P.longifolia seed oil,nitrofurantoin,ciprofloxacin,chloramphenicol,colistin and minocycline than isolates of potentially pathogenic bacteria.The study concludes that HBLs may harbour both good and bacteria,and should be consumed raw only after proper decontamination.

Assessment of the Level of Knowledge of the Nature of Highly Pathogenic Avian Influenza Demonstrated by the Nigerian Veterinary Laboratory Staff Involved in HPAI Diagnosis in Nigeria

The study was designed to evaluate the level of knowledge of Nigerian Veterinary Laboratory Staff on the nature of Highly Pathogenic Avian Influenza (HPAI) disease using structured questionnaires. The study comprised the Staff of National Veterinary Research Institute (NVRI) and five reference Veterinary Teaching Hospitals (VTH) designated for HPAI diagnosis. A total of 69 questionnaires were distributed to the laboratory staff. Questions on the general nature of the disease such as the cause, signs, mode of transmission, methods of identification, lesions, control and prevention, etc. were asked. The results showed that 77.38% of the staff answered all the questions correctly indicating their considerable knowledge of the HPAI disease. Considerable percentage of the staff listed correctly the equipment used for serology (36.23%) and RT-PCR (31.88%). Interestingly only 13.04% of the staff listed correctly the equipment used in rapid tests despite the fact that they are simpler and recommended for all P2 laboratories. In conclusion, the veterinary laboratory staff assessed demonstrated a significant level of knowledge on HPAI diagnosis;however, most of their laboratories lack the structure, organization, funds and basic facilities required for effective HPAI diagnosis.

Immunogenicity of formaldehyde and binary ethylenimine inactivated infectious bursal disease virus in broiler chicks

Infectious bursal disease virus (IBDV) was inactivated by two different chemicals—formaldehyde and binary ethylenimine (BEI). Formaldehyde was used at 0.1% and 0.2%, while BEI was used at concentrations of 0.001 and 0.002 mol/L. These four vaccines were tested for their efficiency in generating humoral immune response in different groups of broiler chicks. Both BEI-inactivated vaccines gave relatively higher antibody titers and were almost twice as efficient as formaldehyde-inactivated ones.

Inactivation of infectious bursal disease virus by binary ethylenimine and formalin

In this experiment conducted to study the inactivation dynamics of infectious bursal disease virus (IBDV) by binary ethylenimine (BEI) in comparison with formalin, IBDV was isolated from the bursa of infected chickens and its confirmation was done by agar gel precipitation test. Viral suspensions were subjected to inactivation with BEI and formalin for pre-set time in-tervals. BEI was employed at concentrations of 0.001 and 0.002 mol/L while formalin was used at 0.1% and 0.2%. Sampling was done at 6, 12, 24, 36 and 48 h of incubation and samples were tested for their inactivation status in 9-day-old embryonated eggs and 3-week-old broiler chickens. IBDV was completely inactivated by 0.001 and 0.002 mol/L BEI after 36 h of incubation at 37 °C, whereas formalin at 0.1% and 0.2% concentrations inactivated IBDV in 24 h.

Staphylococcus aureusβ-hemolysin-neutralizing single-domain antibody isolated from phage display library of Indian desert camel

Objective:To isolate and characterize Staphylococcus aureus(S.aureus)β-hemolysinneutralizing dAbs from phage display library of Indian desert camel.Methods:Phage display library of 5×10 dAb clones of LPS-immunized Indian desert camel constructed in our laboratory was used for selection of S.aureus exotoxin-specific clones by panning technique.Enrichment of Ag-specific clones in successive rounds of panning was assessed by phage-ELISA and phage titration.Different dAb clones binding to S.aureus exotoxin Ags were expressed with C-terminal 6×His tag in E.coli and purified by Ni-chelate chromatography.The expression was verified by SDS-PAGE and western blotting.The purified clones were tested for inhibition of ’hot-cold’ hemolytic activity in vitro.Resistance to thermal inactivation of the dAb clones was studied by observing the effect of heat treatment from 50℃to 99℃for 30 min on the ’hot-cold’ hemolytic activity in vitro.Results:Several dAb clones binding to S.aureus exotoxins were isolated and enriched by three rounds of panning.The soluble dAb clones were approximately～16 kDa in size and reacted with 6×His tag specific murine monoclonal antibody in western blot.One of the Ni-chelate affinity purified dAb.6×His clones,inhibited S.aureusβ-hemolysin activity in vitro and resisted thermal inactivation upto 991.Conclusions:An S.aureusβ-hemolysinneutralizing dAb clone of possible therapeutic potential has been isolated.

Outbreaks of salmonellosis in three different poultry farms of Kerala,India

Avian salmonellosis is an important disease causing serious impediment to the development of poultry industry especially in developing countries of Asia and Africa.Since no"effective"immunoprophylactic measures are available for the disease till date,strict biosecurity is the only alternative to preclude the disease.For formulating the control measures,an understanding of the epidemiology of the disease,proper diagnosis and identification of the causative agent is quintessential.This report sheds light on three different outbreaks of salmonellosis in three different farms in Kerala(India)describing the disease diagnosis,antibiotic resistance and the suggested control measures.All the three isolates were revealed to be Salmonella gallinarum and were resistant to at least three of the antimicrobial agents tested.

Isolation, serotype diversity and antibiogram of Salmonella enterica isolated from different species of poultry in India

Objective: To study the occurrence and serotype diversity of Salmonella isolates in different species of poultry(chicken, emu and duck) and determine their resistance pattern against various antibiotics of different classes.Methods: About 507 samples comprising 202 caecal contents and 305 fecal samples from chicken, emu and duck were processed for isolation of Salmonella enterica. Salmonellae were isolated and detected by standard protocol of ISO 6579 Amendment 1: Annex D. Genetic confirmation was also made by using 16 S r RNA genus specific PCR. Serotype specific PCR was also done to detect the most common serovars viz. Salmonella Enteritidis, Salmonella Typhimurium and Salmonella Gallinarum. All obtained isolates were subjected to a set of 25 antibiotics to study their antibiogram by using Baeur-Kirby disk diffusion method.Results: Out of 507 samples processed, 32 isolates of Salmonella enterica(18 from caecal contents and 14 from faecal samples) were obtained, of which 24 belonged to 6 different serovars, 6 were untypeable and 2 were rough strains. Salmonella Enteritidis was the most predominant serotype(9), followed by Salmonella Typhimurium(5), Salmonella Virchow(4), Salmonella Gallinarum(3), Salmonella Reading(2) and Salmonella Altona(1). Antibiotic resistance pattern was maximum(100%) to oxacillin, penicillin and clindamycin, followed by ampicillin(68.75%), tetracycline(65.62%), nalidixic acid(56.25%) and colistin(46.87%). High sensitivity of isolates was recorded for chloramphenicol(96.87%) followed by meropenem(84.37%). Conclusions: Occurrence of high proportion of serovars in our study which can cause serious gastroenteritis in humans is a matter of concern. Salmonella Altona has been detected for the first time in India from poultry. This serotype is known to cause serious outbreaks of gastroenteritis in humans. Multidrug resistant isolates were recovered at high percentage which can be attributed to non-judicious use of antibiotics both in prophylaxis and treatment regimen. This observation draws serious attention as poultry serves as an important source of transmission of these multidrug resistant Salmonella serovars to humans.

Comparative study of Anaplasma parasites in tick carrying buffaloes and cattle

A comparative study on the prevalence of Anaplasma parasite was conducted on ticks carrying buffaloes and cattle. Five hundred blood samples of both animals (250 of each) were collected during February, March and April. Thin blood smears on glass slides were made, fixed in 100% methyl alcohol and examined. Microscopic examination revealed that 205 (41%) animals had Anaplasma parasites, out of which 89, 44 and 72 animals had Anaplasma marginale, Anaplasma centrale and mixed infection respectively. Infected buffaloes and cattle were 75 and 130 respectively. The infection in female was 53 and 92 in buffaloes and cattle respectively. Twenty-two and 92 blood samples of male were found positive in buffaloes and cattle respectively. Com- parative study revealed that the cattle were 26.82% more susceptible than buffaloes. The parasite prevailing percentage in female of both animals was slightly higher than that of the male. This investigation was aimed at studying the comparative prevalence of Anaplasma parasite in tick carrying buffaloes and cattle.

Mortality rates from a Nigerian isolate of the <i>Infectious Bursa Disease Virus</i>and passive haemagglutination antibody titer that protects chicks against challenge with the virus isolate

To determine passive haemagglutination (PHA) antibody titer that would protect chicks against Nigerian isolates of the Infectious Bursa Disease Virus (IBDV), five groups of chicks aged 30 days which had different antibody titers were challenged with a Nigerian isolate of virulent IBDV. Mortality rates of the different groups were plotted against their respective mean PHA antibody titers. A group with zero antibody titer had a mortality rate of 75% while those with PHA antibody titers of 185.6, 243.2, 256 and 307.2 had mortality rates of 40%, zero, zero and zero respectively. Linear equation generated for a line of best fit of the graph of mortality rates of the chicks on their IBD antibody titers gave antibody titer (X) at which mortality (Y) would be zero as 300. A mortality of 75% and the high antibody level needed to protect chicks suggest that the isolate may be a hypervirulent strain.

Production of spike and nucleocapsid recombinant proteins of porcine epidemic diarrhea virus for antibody detection by ELISA

Porcine epidemic diarrhea (PED), a devastating enteric disease in pigs, is caused by PEDvirus (PEDV)(1)Reduced severity of clinical diseases was reported to associate with neutralizing antibody titers in colostrum. However, viral neutralization assay(VN) is laborious and not suitable for routine diagnosis. Spike protein plays an important role in stimulating neutralizing antibody that might be suitable for PEDV diagnosis.

Modification of the passive heamagglutination test for detection of <i>infectious bursal disease virus</i>

To modify the Passive Haemagglutination (PHA) test, a rapid test, used for qauntitative detection of viral antibodies, so that it can be used for determination of viral titres, dilutions of Infectious Bursal Disease Virus (IBDV) were used to sensitize the Red Blood Cells (RBCs) before reacting them with known IBD serum. Also, to improve sensitivity of the test, different RBC concentrations were used for the test. A standard IBDV gave positive PHA reaction upto its 1:2048 dilution. With different IBDV samples, positive PHA reactions occured upto dilutions, ranging from 1:16 to 1:4096. Different RBC concentrations gave different titres for same IBDV samples. With 0.6% and 0.2% RBC concentrations, mean PHA titres of IBDV samples increased from 454. 85 ± 315.32 to 2396.57 ± 489.55 (p < 0.05 ). It was concluded that PHA can be adopted for evaluation of viral titres. To improve sensitivity of the test, use of 0.2% RBC is recommended.

Fungal Contaminants of Smoke-Dried Fish Sold in Open Markets in Makurdi, Benue State, North-Central Nigeria

This mycological study was embarked upon to provide more detailed reports on the presence and different types of fungal agents associated with the contamination of smoke-dried fish sold in open markets in Makurdi, Benue State. A total of 100 randomly selected smoke-dried fish samples from the 3 major fish markets in the study area were used for the study. The samples were aseptically processed and cultured for fungal isolation using Sabouraud Dextrose Agar. Isolates were identified using macroscopic and microscopic characteristics, and comparison with species in fungal atlas. Fungi were detected in 74 of the 100 smoke-dried fish samples from the 3 markets surveyed with isolation rates ranging from 67.6% to 84.8%. A total of 77 fungal isolates were recorded from the 74 positive samples. Aspergillus (28.6%), Mucor (20.8%), Penicillum (18.2%) and Rhizopus (15.6%) were the dominant fungi detected, while Absidia (9.1%) and Candida (7.8%) occurred less frequently. In view of the high fungal contamination of smoke-dried fish, fishermen and marketers should adapt better methods of fish handling and preservation. More awareness on the health implications of fungal contamination of fish products should be created among fish handlers and consumers.

Immunostimulatory Effect of Levamisole on the Immune Response of Goats to <i>Peste des petits ruminants </i>(PPR) Vaccination

The use of Levamisole hydrochloride as an immunostimulant has been successful in the control of certain diseases such as Derzsy’s disease in chicken as well as in increasing lymphocytes and weight gain in pigs. However, its use as immunostimulant in the prevention of Peste des petits ruminants (PPR) in goats has not been investigated. In this study, the use of Levamisole in enhancing immune response to PPR vaccination including its effects on weight gain was investigated among groups of goats (groups A, B, C and D). Virus neutralization test was used to determine the antibody profile in vaccinated goats while the total and differential leucocyte counts and body mass index (BMI) were determined using standard procedures. Goats in group A (Levamisole primed and PPR vaccinated) seroconverted to more than 3-folds of the initial pre-vaccination geometric mean titre (GMT) of neutralizing antibody beginning from second week post-vaccination and remained high throughout the period of the experiment. Similarly, there was significant increase (p 0.05) in BMI of animals in other groups throughout the period of the experiment. The results of this study further confirm the immunostimulatory effect of Levamisole when used in combination with a vaccine.

FOXO3a as a sensor of unilateral nerve injury in sensory neurons ipsilateral, contralateral and remote to injury

Emerging evidence supports that the stress response to peripheral nerve injury extends beyond the injured neuron,with alterations in associated transcription factors detected both locally and remote to the lesion.Stress-induced nuclear translocation of the transcription factor forkhead class box O3a(FOXO3a)was initially linked to activation of apoptotic genes in many neuronal subtypes.However,a more complex role of FOXO3a has been suggested in the injury response of sensory neurons,with the injured neuron expressing less FOXO3a.To elucidate this response and test whether non-injured sensory neurons also alter FOXO3a expression,the temporal impact of chronic unilateral L4–6 spinal nerve transection on FOXO3a expression and nuclear localization in adult rat dorsal root ganglion neurons ipsilateral,contralateral or remote to injury relative to na?ve controls was examined.In na?ve neurons,high cytoplasmic and nuclear levels of FOXO3a colocalized with calcitonin gene related peptide,a marker of the nociceptive subpopulation.One hour post-injury,an acute increase in nuclear FOXO3a in small size injured neurons occurred followed by a significant decrease after 1,2 and 4 days,with levels increasing toward pre-injury levels by 1 week post-injury.A more robust biphasic response to the injury was observed in uninjured neurons contralateral to and those remote to injury.Nuclear levels of FOXO3a peaked at 1 day,decreased by 4 days,then increased by 1 week post-injury,a response mirrored in C4 dorsal root ganglion neurons remote to injury.This altered expression contralateral and remote to injury supports that spinal nerve damage has broader systemic impacts,a response we recently reported for another stress transcription factor,Luman/CREB3.The early decreased expression and nuclear localization of FOXO3a in the injured neuron implicate these changes in the cell body response to injury that may be protective.Finally,the broader systemic changes support the existence of stress/injury-induced humeral factor(s)influencing transcriptional and potentially behavioral changes in uninjured dorsal root ganglion neurons.Approval to conduct this study was obtained from the University of Saskatchewan Animal Research Ethics Board(protocol#19920164).

Clinical and hematological study of canine Ehrlichiosis with other hemoprotozoan parasites in Kolkata,West Bengal,India

Objective:To observe other hemoprotozoan diseases with canine ehrlichiosis und to evaluate the clinical and hematological aspects of dogs naturally infected with ehrlichiosis with other hemoprotozoun diseases.Methods:Blood was collected for hematological value and Giemsa stained blood smear was made for diagnosis of Ehrlichia sp.and other hemoprotozoan parasites from naturally infected dogs.Case history was taken from the owner and clinical signs and symptoms were noted.Results:A total of 47 cases of ehrlichiosis in dogs were reported with babesiosis(8.51%)and hepatozoonosis(6.38%)hemoprotozoan diseases.Ehrlichia cams,Ehrlichia ewingii,Brucella canis,Babesia gibsoni and Hepatozoon canis were observed under oil immersion lense of microscope in Giemsa stained peripheral blood smears.Marked anaemia and neutrophilic leukocytosis were observed.Conclusions:The results of this study stated that clinical and haeinatological changes occurred in canine ehrlichiosis with babesiosis and hepatozoonosis due to parasitemia.In mixed infection,the disease more severe,and also it depended on immunity of animals.Babesia gibsoni and Hepatozoon canis with Ehrlichia sp.were first reported from West Bengal state of India by this study.

Physico-chemical indices of breeding sites of Simulium damnosum in the lower Cross River Basin, Nigeria

To determine the factor influencing the abundance development and distribution of Simulium damnosum(S.I) immature stages in the lower Cross River Basin, Nigeria, the physico-chemical characteristics of water at breeding sites were analysed longitudinally over a 12 month period. Analysis of the water samples showed that water current correlated positively and significantly(p<0.05), with dissolved oxygen(r = 0.95 vs 0.10); conductivity(r = 0.95 vs 0.81); hydrogen ion concentration(r = 0.03 vs 0.46); and biochemical oxygen demand(r = 0.80 vs 0.64); from Agbokim Waterfalls and Afi River respectively. The result indicates that these parameters and low ionic concentration are the most essential requirement for the development of S. damnosum pre-imaginal stages. Amplitudes of annual variability as measured by co-efficient of variation varied between the parameters. From the trends in hydrological variables it was deduced that the integrated impact of precipitation, input of surface runoff and municipal effluents played an overriding role in determining the absolute levels and temporal pattern in the water quality attributes. Of the 3578 pre-imaginal stages of S. damnosum collected, 52.52% and 47.4% were collected from Agbokim Waterfall and Afi River respectively. There was a significant difference(p<0.05) in the number of pre-imaginal stages collected from both sites. The highest number of pre_imaginal stages of S. damnosum collected were at the peak of rain(July—September) indicating that S. damnosum in the River Basin is a wet season breeder. The need for long term laboratory colonization of blackfly immature stages becomes imperative especially how these parameters could be exploited in control programme through the testing of larvicides without adverse ecological damage to the aquatic habitat.

A looming twindemic of COVID-19 and dengue on post-flood scenario in the developing countries

Pakistan has recently faced a disastrous and lethal flooding across the country,affecting 33 million people and 1400 healthcare facilities,putting further strain on the nation's fragile healthcare system.The country faces widespread infectious diseases including dengue and COVID-19[1].To date,SARS-CoV-2 has caused about 6.53 million deaths and spread globally,infecting about 612 million people.Throughout the developing world,infection rate is particularly high in Pakistan,causing 30606 deaths so far[2].As of September 2022,healthcare authorities claimed a continued rise in dengue cases due to heavy flood in the country.Till date,a total of 14173 dengue cases have been reported,and the highest number of cases was documented in Sindh province(5203),followed by Khyber Pakhtunkhwa(4538),Punjab(3101),and Islamabad with 1331[3].

Profile of Maternally Derived Antibody in Broiler Chicks and In-Ovo Vaccination of Chick Embryo against Newcastle Disease

In this study, we determined the suitability of lentogenic LaSota and naturally occurring avirulent I2 vaccine strains of Newcastle disease (ND) virus for efficacious in-ovo vaccination of broiler chickens. A total of 114 embyonated eggs divided into five groups (A, B, C, D and E) consisting of 25 eggs in each of groups A and B, 20 eggs in each of groups C and D and 24 eggs in group E were used in the study. Eighteen-day-old embryonated eggs in group A were vaccinated in-ovo with ND-I2 vaccine while the same age of embryos in group B were vaccinated with ND-LaSota. Thirteen-day-old embryonated eggs in groups C and D were vaccinated with ND-I2 and ND-LaSota respectively. Group E served as unvaccinated control. There was significant difference (p 2) used for the in-ovo vaccination were pathogenic for chick embryos, however, ND-I2 vaccine was better tolerated when administered to 18-day-old chick embryo.