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This study was conducted to determine the impact of adulteration on physico-chemical and coliforms content of marketed raw milk. For this purpose, 300 milk samples were collected from urban and peri-urban localities of Jhang, Pakistan. The results showed that 13.3%, 9.7%, 9%, 8.3% and 5.7% of the tested milk samples were positive for cane sugar, formalin, boric acid, H2O2 and urea, respectively besides the highest adulteration of water (95%). Other adulterants included soap (4.7%), sodium chloride (3.7%) and quaternary ammonium compounds (3.3%). The incidence of cane sugar and starch adulteration was highest in winter while formalin and H2O2 in summer season. The compositional quality of milk was also deteriorated by relatively lowered concentrations of lactose, fats, total solids, solids-not-fat (SNF) and proteins. Likewise, the pH and freezing point of contaminated milk samples were comparatively higher than the corresponding standard values. Furthermore, the adulterated milk samples were highly contaminated with E. coli (42.5%) and Enterobacter aerogenes (27.5%) as compared to non-adulterated milk. It was concluded that milk adulterationin the study area predominates round the year badly deteriorating the physico-chemical characteristic of milk besides contaminating it with injurious adulterants and water-borne pathogens posing potential health risks to the consumers.

Amino acids play crucial roles in the MTOR (mechanistic target of rapamycin kinase) complex 1 (MTORC1) pathway. However, the underlying mechanisms are not fully understood. Here, we establish a cell-free system to mimic the activation of MTORC1, by which we identify CANX (calnexin) as an essential regulator for leucine-stimulated MTORC1 pathway. CANX translocates to lysosomes after leucine deprivation, and its loss of function renders either the MTORC1 activity or the lysosomal translocation of MTOR insensitive to leucine deprivation. We further find that CANX binds to LAMP2 (lysosomal associated membrane protein 2), and LAMP2 is required for leucine deprivation-induced CANX interaction with the Ragulator to inhibit Ragulator activity toward RRAG GTPases. Moreover, leucine deprivation promotes the lysine (K) 525 crotonylation of CANX, which is another essential condition for the lysosomal translocation of CANX. Finally, we find that KAT7 (lysine acetyltransferase 7) mediates the K525 crotonylation of CANX. Loss of KAT7 renders the MTORC1 insensitivity to leucine deprivation. Our findings provide new insights for the regulatory mechanism of the leucine-stimulated MTORC1 pathway. Abbreviations: CALR: calreticulin; CANX: calnexin; CLF: crude lysosome fraction; EIF4EBP1: eukaryotic translation initiation factor 4E binding protein 1; ER: endoplasmic reticulum; GST: glutathione S-transferase; HA: hemagglutinin; HEK293T: human embryonic kidney-293T; KAT7: lysine acetyltransferase 7; Kcr; lysine crotonylation; KO: knockout; LAMP2: lysosomal associated membrane protein 2; LAMTOR/Ragulator: late endosomal/lysosomal adaptor: MAPK and MTOR activator; MAP1LC3B: microtubule associated protein 1 light chain 3 beta; MTOR: mechanistic target of rapamycin kinase; PDI: protein disulfide isomerase; PTM: post-translational modification; RPS6KB1/p70S6 kinase 1: ribosomal protein S6 kinase B1; RPTOR: regulatory associated protein of MTOR complex 1; SESN2: sestrin 2; TMEM192: transmembrane protein 192; ULK1: unc-51 like autophagy activating kinase 1.

Background The interaction between nutrition and immunity plays a vital role in nutrient digestion, absorption, and metabolism during poultry production. Recent studies showed that the gut microbiota contributes to the development of intestinal mucosal immunity. However, the mechanisms by which gut microbes regulate this process remain unclear. Methods We compared the intestinal mucosal immunity and gut microbiota of Arbor Acre broilers (AA (lower mucosal immunity) and Chinese native Wuliang Mountain Black-bone chickens (WLMB) (higher mucosal immunity) using 16S rDNA sequencing, transcriptomic analysis, and immunoglobulin A (IgA) antibody repertoire sequencing. We then combined 16S rDNA sequencing with transcriptomics to identify the key microbes and found that they were positively correlated with IgA production. Next, we transplanted candidate microbes into 1-day-old broiler to explore their role in intestinal mucosal immunity. Finally, we verified the function of candidate microbial metabolites in regulating the immune function of macrophages and the intestinal-epithelial cells (IECs) using in vitro experiments. Results WLMB performs stronger mucosal immunity than AA, including higher IgA levels, more diverse IgA antibody repertoire, and higher bacterial affinity. Bacteroides was identified as the key microbes related to the intestinal IgA response. Bacteroides transplantation could increase IgA concentration in the duodenal contents by enhancing the expression of IgA, polymeric immunoglobin receptor (PIgR), B cell-activating factor of the TNF family (BAFF), and activation-induced cytidine deaminase (AID) in the duodenum. Additionally, Bacteroides-derived isovaleric acid promoted M2 macrophage polarization of macrophage via mTOR/PPAR-γ/STAT3 signaling pathways and regulated the immunologic function of IECs to produce cytokines, including interleukin (IL)-10, IL-4, BAFF, and transforming growth factor-beta (TGF-β), thus promoting IgA production in B cells by facilitating AID expression. Conclusion Our study revealed that Bacteroides modulate the intestinal IgA response and maintain gut health in broilers. Bacteroides may be a promising alternative as an immunomodulatory microbial agent for developing next-generation probiotics for broiler production.

Inflammation can cause various chronic diseases like inflammatory bowel diseases. Various food protein-derived bioactive peptides (BAPs) with anti-inflammatory activity have the potential to manage these diseases. The aim of this paper is to overview the mechanisms and the molecular targets of BAPs to exert anti-inflammatory activity. In this review, the in vitro and in vivo effects of BAPs on intestinal inflammation are highlighted. The mechanism, pathways, and future perspectives of BAPs as the potential sources of therapeutic treatments to alleviate intestinal inflammation are provided, including nuclear factor-κB, mitogen-activated protein kinase, Janus kinase-signal transducer and activator of transcription, and peptide transporter 1 (PepT1), finding that PepT1 and gut microbiota are the promising targets for BAPs to alleviate the intestinal inflammation. This review provides a comprehensive understanding of the role of dietary BAPs in attenuating inflammation and gives a novel direction in nutraceuticals for people or animals with intestinal inflammation.

Hemorrhagic septicemia is a fatal disease of cattle and buffalo all over the world including Pakistan and it causes heavy economic losses every year. The poor farmers cannot bear this loss in the form of less milk production and heavy expenditures on the animal treatment. An enriched alum-precipitated vaccine with reduced dose was prepared and standardized and safety testing of enriched vaccine was performed in Swiss albino mice as well as in natural host. In this experiment, a total of 36 cattle both male and female of different age groups ranging from 4 months to 4 years were used. All these animals belong to all major cattle breeds of Pakistan including Sahiwal, Red Sindhi (crossbred and purebred), Dhani (crossbred and purebred), Lohani (crossbred and purebred), and Cholistani, and exotic breeds including Holstein Friesian and Jersey. These animals were examined for current immune titer prior to vaccination. Animals were vaccinated subcutaneously with 2 mL and 4 mL dose of new vaccine and were observed for any untoward reaction for 48 h. All the animals were kept under close observation for the next 30 days and all were found safe. The experiment was designed to reduce the dose of the vaccine to 2 mL by using BHI as a growth medium, as well as to increase the number of doses prepared in the same infrastructure, hence reducing the cost of vaccine production. The study proved that vaccine with increased biomass in reduced dose is safe in local as well as in exotic breeds of cattle.

The study was aimed to investigate the influence of animal age, post-slaughter chilling rate, and aging time on meat quality of M. longissimus dorsi (LD) of water buffalo (Bubalus bubalis) and humped cattle (Bos taurus indicus) bulls. After slaughtering, one side of carcasses was subjected to rapid chilling (RC) (0 ± 2°C) and other side was hanged in controlled room temperature (25 ± 2°C) for 3 hr, then allowed to the chiller (0 ± 2°C). The meat quality traits were analyzed at 1, 7, and 14 days of storage. It was noted that rapidly chilled carcasses from the younger animals of both species missed the ideal pH/temperature window, which affects the toughness of the meat. Buffalo meat presented higher shear force, color L* values, and lower b* value as compared to the cattle meat. Moreover, meat shear force values decreased while all color coordinates and cooking loss values increased with lengthening the storage time in both age groups of cattle and buffalo. In conclusion, the tenderness of cattle meat was superior to that of buffalo and RC adversely affect the shear force values of young cattle and both age groups of buffalo bulls.