Resumo O presente estudo envolve a análise química e bacteriológica de água de diferentes fontes, ou seja, furo, poços, garrafa e torneira, dos distritos de Peshawar, Mardan, Swat e Kohat da província de Khyber Pakhtunkhwa (KP), Paquistão. De cada distrito, 50 amostras de água (10 amostras de cada fonte), independentemente do status urbano e rural, foram coletadas dessas fontes e analisadas para sulfatos, nitratos, nitritos, cloretos, sólidos solúveis totais e coliformes (E. coli). Os resultados indicaram que a maioria das fontes de água tinha uma contagem inaceitável de E. coli, ou seja, > 34 UFC / 100 mL. As amostras positivas para E. coli foram elevadas no distrito de Mardan, seguido por Kohat, Swat e distrito de Peshawar. Além disso, algumas fontes de água também foram contaminadas quimicamente por diferentes fertilizantes inorgânicos (nitratos/nitritos de sódio, potássio), mas em níveis seguros, enquanto os resíduos agrícolas e industriais (compostos de cloreto e sulfato) estavam em níveis inseguros. Entre todos os distritos, a qualidade da água foi considerada comparativamente mais deteriorada nos distritos de Kohat e Mardan do que nos distritos de Peshawar e Swat. Essas fontes de água química e bacteriologicamente impróprias para beber podem causar problemas à saúde humana.

Abstract The present study involves the chemical and bacteriological analysis of water from different sources i.e., bore, wells, bottle, and tap, from Peshawar, Mardan, Swat and Kohat districts of Khyber Pakhtunkhwa (KP) province, Pakistan. From each district, 50 water samples (10 samples from each source), regardless of urban and rural status, were collected from these sources and analysed for sulphates, nitrates, nitrites, chlorides, total soluble solids and coliforms (E. coli). Results indicated that majority of the water sources had unacceptable E. coli count i.e.> 34 CFU/100mL. E. coli positive samples were high in Mardan District, followed by Kohat, Swat and Peshawar district. Besides this, the some water sources were also chemically contaminated by different inorganic fertilizers (nitrates/nitrites of sodium, potassium) but under safe levels whereas agricultural and industrial wastes (chloride and sulphate compounds) were in unsafe range. Among all districts, the water quality was found comparatively more deteriorated in Kohat and Mardan districts than Peshawar and Swat districts. Such chemically and bacteriologically unfit water sources for drinking and can cause human health problems.

Resumo Os lactobacilos são probióticos com capacidade de desintoxicação da Aflatoxina (AF), encontrados em produtos fermentados, TGI de animais e meio ambiente. O objetivo deste estudo foi investigar a capacidade de isolados de frango de corte de Lactobacillus contra a Aflatoxina B1 (AFB1). Para tanto, 5 isolados de Lactobacillus de intestino de frango foram incubados com 100 ppb AFB1 em meio aquoso, e o efeito de diferentes parâmetros (frações celulares, tempo, temperatura, pH) na desintoxicação foi determinado por CLAE. O efeito melhorador de Lactobacillus salivarius (LS) contra AFB1 foi estudado em frangos de corte. Os resultados revelaram que LS (CR. 4) apresentou os melhores resultados (in vitro) em comparação com outros isolados [L. salivarius (CR. 3, CR. 4), L. agilis (CE. 2.1, CE. 3.1) e L. crispatus (CE. 28)]. Detritos celulares de CR. 4 mostraram desintoxicação significativamente maior (P < 0.05). A quantidade máxima de AFB1 foi desintoxicada a 30 °C (97%), pH 4.0 (99%) e 6 h (99,97%). O estudo in vivo mostrou que AFB1 diminuiu o ganho de peso (1,269 ± 0.04 g / ave), alimento consumido (2,161 ± 0.08 g / ave), proteína total sérica (2.42 ± 0.34 g / dl), albumina sérica (0.5 ± 0.22 gm / dl) e título de anticorpo (4.2 ± 0.83). As enzimas da função hepática foram encontradas (alanina transaminase (ALT): 32 ± 10.7 U / L) e aspartato transaminase (AST): 314.8 ± 27 U / L) elevadas em AFB1 alimentados com frangos. O tratamento com 1% LS não só diminuiu os efeitos tóxicos de AFB1 (grupo D), mas também melhorou a saúde geral dos frangos devido aos seus efeitos probióticos (p < 0.05) em comparação com o controle negativo (grupo A). A capacidade de desintoxicação do LS foi melhor do que o aglutinante comercial (CB) (0.2% Protmyc). Concluiu-se que a desintoxicação de AFB1 por Lactobacillus foi dependente da cepa, temperatura, pH e tempo. LS tem capacidade de desintoxicação contra AFB1 in vivo.

Abstract Lactobacilli are probiotics with Aflatoxin (AF) detoxification ability, found in fermented products, GIT of animals and environment. Purpose of this study was to investigate the ability of broiler isolates of Lactobacillus against Aflatoxin B1 (AFB1). For this purpose, 5 isolates of Lactobacillus from broiler gut were incubated with 100 ppb AFB1 in aqueous environment and effect of different parameters (cell fractions, time, temperature, pH) on detoxification was determined by HPLC. The ameliorative effect of Lactobacillus salivarius (LS) against AFB1 was studied in broiler. The results revealed that LS (CR. 4) showed the best results (in vitro) as compared to other isolates (L. salivarius (CR. 3, CR, 4), L. agilis (CE. 2.1, CE. 3.1) and L. crispatus (CE. 28). Cell debris of CR. 4 showed significantly higher detoxification (P<0.05). Maximum amount of AFB1 was detoxified at 30°C (97%), pH 4.0 (99%) and 6 h (99.97%). In vivo study showed that AFB1 decreased weight gain (1,269 ± 0.04 gm/ bird), feed consumed (2,161 ± 0.08 gm/ bird), serum total protein (2.42 ± 0.34 gm/ dl), serum albumin (0.5 ± 0.2 2 gm/dl) and antibody titer (4.2 ± 0.83). Liver function enzymes were found (alanine transaminase (ALT): 32 ± 10.7 U/L) and aspartate transaminase (AST): 314.8 ± 27 U/L) elevated in AFB1 fed broilers. Treatment with 1% LS not only decreased the toxic effects of AFB1 (group D) but also improved the overall health of broilers due to its probiotic effects (p<0.05) as compared to control negative (group A). The detoxification ability of LS was better than commercial binder (CB) (0.2% Protmyc). It was concluded that detoxification of AFB1 by Lactobacillus was strain, temperature, pH and time dependent. LS has detoxification ability against AFB1 in vivo.

ABSTRACT This study aimed to assess the impact of incorporating Citrullus colocynthis fruit pulp (CCFP) into the diet of broiler chickens as a replacement for antibiotic growth promoters (AGPs) on growth performance, blood biochemistry, and intestinal microbial count. A total of 600 unsexed Ross-308 day-old broiler chicks were randomly assigned to 6 groups using a completely randomized design. Each group had 5 replicates, with 20 chicks per replicate. The study included a control group (T1) that did not receive any CCFP or AGP supplementation. The other experimental groups were as follows: T2 (0.15 g/kg AGP, 0 g/kg CCFP), T3 (0.11 g/kg AGP, 0.75 g/kg CCFP), T4 (0.075 g/kg AGP, 1.5 g/kg CCFP), T5 (0.037 g/kg AGP, 2.25 g/kg CCFP), and T6 (0 g/kg AGP, 3 g/kg CCFP). The results indicated a reduction in feed intake (p = 0.045) in group T3 compared to the control group (T1). Blood biochemical analysis showed that group T6 had higher levels of serum total protein (p = 0.050) and globulin (p = 0.044) compared to the control group (T1). The cecal microbial count revealed a lower total bacterial count (p = 0.01) in groups T2 and T3, and a reduced E. coli count (p = 0.050) in group T3 compared to the group T5. However, the ileal microbial count showed no significant differences between treatments. In conclusion, these findings suggest that a diet containing 0.75 g/kg CCFP combined with 0.11 g/kg AGP may improve the growth of broiler chickens while maintaining a healthy microbial balance. (CCFP AGPs (AGPs performance biochemistry 60 Ross308 Ross 308 Ross-30 dayold day old design replicates 2 replicate T1 T (T1 supplementation follows 0.15 015 15 (0.1 gkg g kg CCFP, , CCFP) 011 11 075 75 0.7 0.075 0075 (0.07 1 1. 0.037 0037 037 (0.03 225 25 2.2 ( CCFP. . p 0.045 0045 045 T1. 0.050 0050 050 0.044 0044 044 0.01 001 01 E However treatments conclusion 0.1 balance Ross30 30 Ross-3 (T (0. 07 7 0. 0.07 007 (0.0 0.03 003 03 22 2. 0.04 004 04 0.05 005 05 0.0 00 Ross3 Ross-

ABSTRACT The present study intended to identify the effects of the Saccharomyces cerevisiae yeast on production performance and parameters related to the quality of layers’ eggs. Effects of yeast on immune titers after routine vaccinations against Newcastle disease and infectious bronchitis were also studied. Fayoumi chicken (n=288) were divided in four groups (72 in each), and Black Australorp (n=288) in four groups (72 in each). Both had 12 weeks of age and were kept in 08 groups (3 replicates per treatment). Hens were fed a controlled diet along with the addition of 0.5g, 1.0 g, and 1.5 g of S. cerevisiae per kg of feed, till 25 weeks of age. Feed intake and egg production were measured on a daily basis, while body weight gain and egg weight were measured weekly. Egg quality parameters were evaluated by picking 5 eggs from every group weekly. Birds were vaccinated for Newcastle disease (ND) and infectious bronchitis (IB) at the 18th week. Subsequently, 10 days after vaccination, antibody titers were determined by Hemagglutination Inhibition and Enzyme Linked Immunosorbent Assay for both diseases. Results showed no significant effect of S. cerevisiae on layers’ weight gain and feed intake. However, egg production was increased in the experimental group. Moreover, yeast supplementation impacted positively on birds’ immune system. In conclusion, probiotic supplementation improved birds’ egg production and immunity. layers studied n=288 n288 n 288 (n=288 72 (7 each, each , each) each. . 1 0 3 ( treatment. treatment treatment) 05g 5g 0.5g 1. 15 S 2 basis weekly ND (ND IB (IB th week Subsequently vaccination diseases However Moreover birds system conclusion immunity n=28 n28 28 (n=28 7 n=2 n2 (n=2 n= (n= (n

RESUMO Trichostrongylid em ovelhas foi estudado no Distrito Zhob, Balochistan. Foram coletados 120 tratos gastrointestinais (GIT) de ovelhas do matadouro do distrito. Estas amostras foram processadas para isolamento e identificação de nematódeos tricostrônquicos no Laboratório do Hospital Veterinário do Distrito Zhob. A taxa de prevalência geral foi de 39,1% em machos e 60,8% em fêmeas (p=0,001). A taxa de prevalência em duas raças viz Balochi e Rakhshani foi de 58,3% e 41,6%, respectivamente (p=0,01). A prevalência da espécie observada com Trichostrongylus foi 19,1%, Haemonchus foi 20,8%, Cooperia foi 29,1% e Nematodirus foi 30,8% (p=0,087). Quanto a quantidade de espécies infestadas pelas ovelhas, um único tipo de parasita estava em 32,5% de animais, dois tipos de espécies parasitárias em 36,3% dos animais e três tipos de espécies parasitárias em 30,8% dos animais (p=0,366).

RESUMO: O presente estudo foi realizado para verificar o potencial antifúngico do extrato de caule de Sisymbrium irio L. contra Fusarium oxysporum f. sp. cepae (FOC). Em bioensaios preliminares, diferentes concentrações (1% a 5%) de extratos de folhas, caules e frutos foram avaliadas contra o FOC. Todos os extratos foram eficazes contra o patógeno. No entanto, o extrato de folhas foi o mais eficaz, causando 25-41% de redução na biomassa FOC. O fracionamento do extrato metanólico das folhas foi feito por dois solventes orgânicos: n-hexano e clorofórmio. Diferentes concentrações (1,56 a 200 mg mL-1) dessas frações foram testadas contra o FOC. As frações de n-hexano e clorofórmio mostraram atividade inibitória contra o patógeno e resultaram em redução de 77-93% e 80-96% na biomassa de FOC, respectivamente. A análise por CG-EM mostrou a presença de 24 compostos em n-hexano e 4 compostos em fração clorofórmica. Na fração n-hexano, o β-sitosterol (18,64%) foi o composto mais abundante, seguido pelo ácido orótico, éster bis (terc-butildimetilsilil) terc-butildimetilsilílico (12,18%), éster de ácido 10-octadecenoico metil (7,90%) e Ter de di-isoctilo do ido 1,2-benzenodicarboxico (6,05%). Os principais compostos identificados na fração clorofórmica foram o ácido 1,3-benzenodicarboxílico, o éster bis (2-etil-hexílico) (50,82%) e o ftalato di-n-octilo (33,00%). Este estudo conclui que as frações de n-hexano e clorofórmio do extrato metanólico das folhas de S. irio contêm potentes constituintes antifúngicos para o manejo do FOC.

ABSTRACT: The present study was carried out to check the antifungal potential of Sisymbrium irio L. shoot extract against Fusarium oxysporum f. sp. cepae (FOC). In preliminary bioassays, different concentrations (1 to 5%) of leaf, stem and fruit extracts were evaluated against FOC. All the extracts were effective against the pathogen. However, the leaf extract was found the most effective causing 25-41% decrease in FOC biomass. The fractionation of methanolic leaf extract was done by two organic solvents namely n-hexane and chloroform. Different concentrations (1.56 to 200 mg mL-1) of these fractions were tested against FOC. The n-hexane and chloroform fractions showed inhibitory activity against the pathogen and resulted in 77-93% and 80-96% reduction in biomass of FOC, respectively. GC-MS analysis showed the presence of 24 compounds in n-hexane and 4 compounds in chloroform fraction. In n-hexane fraction, β-sitosterol (18.64%) was the most abundant compound followed by orotic acid, bis(tert-butyldimethylsilyl)-, tert-butyldimethylsilyl ester (12.18%), 10-octadecenoic acid, methyl ester (7.90%) and 1,2-benzenedicarboxylic acid, diisooctyl ester (6.05%). Major compounds identified in chloroform fraction were 1,3-benzenedicarboxylic acid, bis(2-ethylhexyl) ester (50.82%) and di-n-octyl phthalate (33.00%). This study concludes that n-hexane and chloroform fractions of methanolic leaf extract of S. irio contain potent antifungal constituents for the management of FOC.

RESUMO: A cebola é atacada por um patógeno fúngico no solo, Fusarium oxysporum, que resulta na doença da podridão basal. No presente estudo, três espécies de Trichoderma (T. pseudokoningii, T. harzianum e T. reesei) e as folhas da planta daninha solanácea Withania somnifera foram utilizadas para o manejo dessa doença. O estudo de interação in vitro revelou que T. harzianum é o agente de biocontrole mais eficaz contra o patógeno. Em um ensaio em vaso, misturou-se material de folha seca de W. somnifera (1%, 2% e 3% p/p) e o inóculo de T. harzianum no substrato já inoculado com o patógeno. A maior incidência da doença (87%) foi observada em controle positivo (inoculação de patógenos sem qualquer alteração). Diferentes doses do material da folha seca reduziram a incidência da doença para 41-66%. Em combinação com o material foliar, T. harzianum reduziu a incidência da doença para 20-53%. Em um bioensaio laboratorial, foi preparado um extrato de folhas secas de W. somnifera em metanol e particionado com n-hexano, clorofórmio, acetato de etilo e n-butanol. A concentração mais elevada (200 mg mL-1) de todas as frações, exceto a de n-butanol, diminuiu significativamente a biomassa fúngica em relação ao controle. Este estudo conclui que a podridão basal da cebola pode ser controlada pela aplicação combinada de material de folha seca de W. somnifera e o agente de controle biológico T. harzianum.

ABSTRACT: Onion is attacked by destructive soil-borne fungal plant pathogen Fusarium oxysporum f. sp. cepae, resulting in basal rot disease. In the present study, three Trichoderma species (T. pseudokoningii, T. harzianum and T. reesei) and leaves of solanaceous weed Withania somnifera were used for management of this disease. The in vitro interaction study revealed T. harzianum as the most effective biocontrol agent against the pathogen. In a pot trial, dried leaf material of W. somnifera (1%, 2% and 3% w/w) and inoculum of T. harzianum were mixed in the pot soil previously inoculated with the pathogen. The highest incidence of the disease (87%) was found in positive control (pathogen inoculation without any amendment). Different rates of dry leaf material reduced the incidence of the disease to 41-66%. T. harzianum in combination with leaf material reduced the incidence of the disease to 20-53%. In a laboratory bioassay, the dry leaf extract of W. somnifera was prepared in methanol and partitioned with n-hexane, chloroform, ethyl acetate and n-butanol. The highest concentration (200 mg mL-1) of all except for the n-butanol fraction significantly decreased fungal biomass over control. This study concludes that basal rot of onion can be controlled by combined application of W. somnifera dry leaf material and biological control agent T. harzianum.

The presence of heavy metals in various farm inputs, including feed, fertilizer, water and environment leads to excretion of the residues in animals' milk. Because consumption of milk contaminated with heavy metals poses serious threats to consumers' health, a study was conducted in 2012 - 2013 in Pakistan to evaluate the concentration of heavy metals in unprocessed raw milk procured in different seasons from Jhang city, Punjab, Pakistan. A total of 400 milk samples was collected from milk shops and dairy farms, and the contents of heavy metals, i.e. copper (Cu), lead (Pb), cadmium (Cd) and chromium (Cr) were analysed via atomic absorption spectrometry. Results indicated that the average concentrations of Cu, Pb and Cd were significantly higher than International Dairy Federation Standards. Within a season, heavy metal levels differed significantly among samples. For each type of milk source, there was a significant difference in mean concentration of heavy metals between summer and winter. Based on these findings, there is a critical need to set legal limits and surveillance for heavy metals in an animal's milk.