Chronic nasal inflammation early in life induces transient and long-term dysbiosis of gut microbiota in mice.

The gut microbiota begins to colonize the host body following birth, develops during the suckling period and changes to the adult type after weaning. The early gut microbiota during the suckling period is thought to have profound effects on the host physiology throughout life but it is still unclear whether early dysbiosis is retained lifelong. Our previous study indicated that chronic nasal inflammation induces dysbiosis of gut microbiota in adult mice. In the present study, we addressed the question as to whether early exposure to chronic nasal inflammation induces dysbiosis, and if so, whether the dysbiosis is retained until adulthood and the sex differences in this effect. Male and female mice received repeated intranasal administration of lipopolysaccharide (LPS) or saline twice a week from P7 to P24 and were weaned at P24. The cecal contents were obtained for 16S rRNA analysis at 2 time points: at 4 weeks (wks), just after weaning, and at maturation to adulthood at 10 wks. The body weight did not differ between saline- and LPS-treated mice till around weaning, suggesting that the mothers' milk was given similarly to all mice. At 4 wks, the beta diversity was significantly different between saline- and LPS-treated male and female mice and the composition of the gut microbiota changed in LPS-treated mice. The abundance of phylum Bacteroidota tended to decrease and that of Firmicutes increased in LPS-treated male mice, while the abundance of Deferribacterota increased in LPS-treated female mice. At 10 wks, the beta diversity was not different between saline- and LPS-treated mice, but the abundance of family Lachnospiraceae significantly decreased in LPS-treated male and female mice by LEfSe analysis. Together, chronic nasal inflammation early in life caused transient and long-term dysbiosis of gut microbiota, which may contribute to the onset and progress of metabolic and neuropsychiatric disorders.

The Role of Milk Oligosaccharides in Enhancing Intestinal Microbiota, Intestinal Integrity, and Immune Function in Pigs: A Comparative Review.

The objective of this review was to identify the characteristics and functional roles of milk coproducts from human, bovine, and porcine sources and their impacts on the intestinal microbiota and intestinal immunity of suckling and nursery pigs. Modern pig production weans piglets at 3 to 4 weeks of age, which is earlier than pigs would naturally be weaned outside of artificial rearing. As a result, the immature intestines of suckling and nursery pigs face many challenges associated with intestinal dysbiosis, which can be caused by weaning stress or the colonization of the intestines by enteric pathogens. Milk oligosaccharides are found in sow milk and function as a prebiotic in the intestines of pigs as they cannot be degraded by mammalian enzymes and are thus utilized by intestinal microbial populations. The consumption of milk oligosaccharides during suckling and through the nursery phase can provide benefits to young pigs by encouraging the proliferation of beneficial microbial populations, preventing pathogen adhesion to enterocytes, and through directly modulating immune responses. Therefore, this review aims to summarize the specific functional components of milk oligosaccharides from human, bovine, and porcine sources, and identify potential strategies to utilize milk oligosaccharides to benefit young pigs through the suckling and nursery periods.

A spike-based mRNA vaccine that induces durable and broad protection against porcine deltacoronavirus in piglets.

Coronaviruses (CoVs) are important pathogens for humans and other vertebrates, causing severe respiratory and intestinal infections that have become a threat to public health because of the potential for interspecies transmission between animals and humans. Therefore, the development of safe, effective vaccines remains a top priority for the control of CoV infection. The unique immunological characteristics of vaccines featuring messenger RNA (mRNA) present an advantageous tool for coronavirus vaccine development. Here, we designed two lipid nanoparticle (LNP)-encapsulated mRNA (mRNA-LNP) vaccines: one encoding full-length spike (S) protein and the other encoding the spike ectodomain (Se) from porcine deltacoronavirus (PDCoV). Fourteen days after primary immunization, both mRNA vaccines induced high levels of immunoglobulin G and neutralizing antibodies in mice, with the S vaccine showing better performance than the Se vaccine. Passive immune protection of the S mRNA vaccine in suckling piglets was confirmed by the induction of robust PDCoV-specific humoral and cellular immune responses. The S mRNA vaccine also showed better protective effects than the inactivated vaccine. Our results suggest that the novel PDCoV-S mRNA-LNP vaccine may have the potential to combat PDCoV infection. IMPORTANCE: As an emerging porcine enteropathogenic coronavirus, porcine deltacoronavirus (PDCoV) has the potential for cross-species transmission, attracting extensive attention. Messenger RNA (mRNA) vaccines are a promising option for combating emerging and re-emerging infectious diseases, as evidenced by the demonstrated efficacy of the COVID-19 mRNA vaccine. Here, we first demonstrated that PDCoV-S mRNA-lipid nanoparticle (LNP) vaccines could induce potent humoral and cellular immune responses in mice. An evaluation of passive immune protection of S mRNA vaccines in suckling piglets confirmed that the protective effect of mRNA vaccine was better than that of inactivated vaccine. This study suggests that the PDCoV-S mRNA-LNP vaccine may serve as a potential and novel vaccine candidate for combating PDCoV infection.

Mitigating effects of selenium and zinc on oxidative stress and biochemical and histopathological changes in lung during prenatal and lactational exposure rats to barium chloride.

Selenium (Se) and zinc (Zn) are essential trace elements with antioxidant properties, and their supplementation has been shown to be protective against the toxicity of various environmental and dietary substances. The aim of this study was to investigate the potential protective effect of selenium and zinc as adjuvants against barium (Ba) toxicity in lactating rats and their offspring. The pregnant rats were divided into six groups: the first as control; group 2 received barium (67 ppm) in the drinking water; group 3 had combined Ba + Se (0.5 mg/kg) in the diet; group 4 received Zn (50 mg/kg bw) by gavage together with Ba; groups 5 and 6, positive controls, were treated with selenium (0.5 mg/kg) and zinc (50 mg/kg bw), respectively. MDA, H2O2, AOPP, CAT, GPx, and SOD levels were measured and lung histopathology was performed. Our results showed that barium administration caused lung damage as evidenced by an increase in MDA, H2O2, and AOPP levels and a decrease in the activities of CAT, GPx, and SOD in mothers and their offspring. A decrease in lung GSH, NPSH, and MT levels was also observed. Supplementation of Ba-treated rats with Se and/or Zn significantly improved the pulmonary antioxidant status of mothers and their offspring. Histopathological examinations were also consistent with the results of biochemical parameters, suggesting the beneficial role of Se and Zn supplementation, as evidenced by less accumulation of collagen fibers as studied by hematoxylin and eosin (H&E) and Masson's trichrome staining. In conclusion, we demonstrate the adverse effects of maternal barium exposure during pregnancy and on neonatal lung health and the protective effects of selenium and zinc in preventing the adverse effects of barium exposure.

Can milk replacer allowance affect animal performance, body development, metabolism, and skeletal muscle hypertrophy in pre-weaned dairy kids?

We aimed to evaluate performance, body development, metabolism, and expression of genes related to skeletal muscle hypertrophy in non-castrated male dairy kids fed with different levels of MR during the pre-weaning period. Sixty newborn male kids, not castrated, from Saanen and Swiss Alpine breeds, with an average body weight (BW) of 3.834 ± 0.612 kg, were distributed in a randomized block design. Breeds were the block factor in the model (random effect). Kids were allocated into 2 nutrition plans (n = 30 kids per treatment) categorized as follows: low nutritional plan (LNP; 1L MR/kid/day) or high nutritional plan (HNP; 2L MR/kid/day). All kids were harvested at 45 d of life. The majority of nitrogen balance variables were affected by the nutritional plan (P < 0.050). Morphometric measures and body condition score (2.99 - LNP vs. 3.28 - HNP) were affected by nutritional plan (P < 0.050), except hip height, thoracic depth and hip width. The nutritional plan affected the body components (P < 0.050), except esophagus and trachea. Animal performance and carcass traits were influenced by nutritional plan (P < 0.050), except carcass dressing (48.56% on average). Nutritional plan affected (P < 0.050) some blood profile variables as the total cholesterol (141.35 vs. 113.25 mg/dL), triglycerides (60.53 vs. 89.05 mg/dL), LDL (79.76 vs. 33.66 g/mL) and IGF-1 (17.77 vs. 38.55 ng/mL) for LNP and HNP respectively. Hypertrophy was greater in HNP than LNP animals (P < 0.050), being represented by the proportion of sarcoplasm (39.76 vs. 31.99%). LNP had a greater mTOR abundance than HNP (P = 0.045), but AMPK was not affected by the nutritional plan. Our findings show that a higher milk replacer allowance enhances animal performance, body development, metabolic parameters, and cellular hypertrophy in pre-weaned dairy kids.

Effect of suckler cow breed type and parity on the development of the cow-calf bond post-partum and calf passive immunity.

BACKGROUND: Development of the cow-calf bond post-partum and passive immunity of calves from spring-calving beef × beef (B×B) and beef × dairy (B×D) cow genotypes was determined using primiparous and multiparous (Experiment 1), and primiparous and second-parity (Experiment 2) animals. In Experiment 1, calves either suckled colostrum naturally ('natural-suckling') (n = 126), or were fed colostrum, using an oesophageal-tube ('artificially-fed') (n = 26), from their dam within 1-h post-partum. In Experiment 2, all calves (n = 60) were artificially-fed colostrum from their dam. Prior to colostrum suckling/feeding, colostrum was sampled for IgG analysis. The cow-calf bond was assessed using CCTV recordings during the first 4-h post-partum. Calves were blood sampled at 48-h post-partum to determine IgG and total protein (TP) concentrations, and zinc sulphate turbidity (ZST) units. RESULTS: There was no difference (P > 0.05) in cow licking behaviours and calf standing and suckling behaviours between the genotypes, except in Experiment 2 where B×D calves had more attempts to suckle before suckling occurred (P ≤ 0.05) compared to B×B calves. In Experiment 1, multiparous cows licked their calves sooner (P ≤ 0.05) and for longer (P < 0.01), and their calves had fewer attempts to stand (P < 0.001), stood for longer (P = 0.05), and had fewer attempts to suckle before suckling occurred (P < 0.001) than primiparous cows; there was no parity effect on cow-calf behaviour in Experiment 2. Colostrum IgG concentrations and measures of calf passive immunity did not differ (P > 0.05) between the genotypes in either Experiment. In Experiment 1, colostrum IgG concentrations were greater (P ≤ 0.05) in multiparous compared to primiparous cows and their calves had superior (P ≤ 0.05) passive immunity; no effect of parity was found in Experiment 2. Passive immunity did not differ (P > 0.05) between suckled and artificially-fed calves in Experiment 1. CONCLUSIONS: Cow genotype had little effect on cow-calf behaviours, but under 'natural-suckling' conditions primiparous cows expressed maternal inexperience and their calves were less vigorous than multiparous cows. Colostrum IgG concentration and calf passive immunity measures were unaffected by genotype, but under 'natural-suckling' conditions calves from primiparous cows had lower passive immunity.

Cow-calf contact: a single-herd observational study of AMS yield during the first 100 days in milk.

An increasing number of dairy farmers plan to implement cow-calf contact (CCC) in their herd which necessitates descriptions of the cows` performance in different systems. The aim of the study was to describe (1) Automatic milking system (AMS) milk yield of cows in a CCC system during the first 100 days in milk (DIM) and (2) AMS milk yield before and after cow-calf separation. In a prospective study at a commercial Norwegian dairy farm, we included all calvings from Norwegian Red cows between January 2019 to April 2020. After calving, cow-calf pairs stayed in an individual calving pen during the first 5-6 d before they were moved to the loose housing unit with the remaining herd. Calves had whole-day (24 h/d) and full physical contact to the cows. Cows were milked in an AMS. From 14 individual cows of which one cow calved twice during the study period, we collected daily AMS yields from 15 different lactations, with parities ranging from 1 (n = 6), 2 (n = 5) and 3 (n = 4). Due to the sample size and structure of the data set, we only calculated descriptive statistics from DIM 7-100. All data is shown separately for primiparous and multiparous cows. Mean (± SD) calf age at (fence-line) separation was 52 d ± 14.8 beyond which suckling was prevented. Our data indicates great individual variation in the AMS milk yield. Prior to separation, primiparous cows` AMS yields ranged from 11.0 to 25.9 kg/d while that of multiparous cows ranged from 4.8 to 28.8 kg/d. Once calves were no longer allowed to suckle, the yield increased gradually. During the week after separation, AMS yields ranged from 17.3 to 30.4 kg/d for primiparous cows and 8.7 to 41.8 kg/d for multiparous cows and these yields increased in DIM 93-100 (26.5 to 34.3 and 20.6 to 38.3 kg/d respectively). This study is limited by a low sample size from a single-herd but may provide useful descriptions of AMS milk yield in a whole-day, full contact CCC system during the first 100 days of lactation.

Supplementing Monosodium Glutamate in Sow Diets Enhances Reproductive Performance in Lactating Sows and Improves the Growth of Suckling Piglets.

In most current farm operations, lactating sows need to overcome reproductive and environmental stresses that have resulted in poor sow production performance and piglet growth. Therefore, this study aimed to investigate the effects of in-feed supplementation of monosodium glutamate (MSG) in sows during late gestation lactation in regard to litter performance. The study subjects were 12 multi-parity sows (Landrace × Large White), farrowing sows with an average parity of four (three with three parities, seven with four parities, and two with five parities). They were randomly divided into the following two diet groups: the basal diet as a control (CON) group based on corn and soybean meal; and the basal diet + 2% MSG group. The experimental time ranged from 109 days before delivery to 21 days after delivery. There were six sows in each group, and each sow served as the experimental unit. There were no significant differences (p > 0.05) in body weight (BW), back fat (BF) thickness and estrus interval between sows supplemented with 2% MSG in their diets before and after farrowing and during weaning (p > 0.05). However, MSG-treated sows tended to increase BW loss at farrowing more than the CON group (p = 0.093) but lost less weight during lactation than the CON group (p = 0.019). There were no significant differences in the body condition scores (BCSs) and BF loss of the two groups of sows before and after farrowing and at weaning (p > 0.05). There was no significant difference in the weight of newborn piglets between the two groups of sows (p > 0.05). The weaning weight (p = 0.020) and average daily gain (ADG) (p = 0.045) of suckling piglets were higher in the MSG treated group compared to the CON group. The daily milk production of sows in the MSG treatment group was higher compared to the CON group (p = 0.045). The protein concentration of milk at week 3 (p = 0.060) and fat concentration of milk at week 5 (p = 0.095) of the MSG-supplemented sows tended to increase more than the CON group. In summary, the dietary inclusion of MSG supplementation had a beneficial effect on the late gestating sows and their piglet's growth and milk production. Our research has shown that the addition of 2% MSG in late gestation and lactation diet would be beneficial for both sow and piglet production.

Boric Acid in Milk Replacer as a Health Enhancer and Growth Promoter for Lambs in the Suckling Period.

This study was performed to investigate the effects of boric acid supplementation in milk replacer of lambs in the suckling period on performance, biochemical parameters, the antioxidant system, fecal culture, and expression of some genes. During the suckling period, 60 lambs (4 days old) were randomly given four levels of boric acid (0, 30, 60, and 90 mg/kg body weight) via milk replacer for 57 days. The lambs supplemented with boric acid had a higher weight gain and better feed conversion ratio. Boric acid supplementation quadratically increased serum triglyceride, total protein, alkaline phosphatase, serum antioxidant activity and oxidative stress biomarkers, and fecal flora and decreased IL1ß, IL10, iNOS, NF-kB, and TNF-α gene expressions. The effect of boric acid on rumen papilla development could not be determined since the animals were not slaughtered. In conclusion, the use of boric acid to lambs in the suckling period improved the average weekly body weight gain and feed conversion efficiency, positively affected some biochemical parameters, antioxidant system, and intestinal flora, and also affected gene expressions related to the immune system. Boric acid supplementation had a beneficial effect on the health and growth of suckling lambs.

The effects of exposure to and timing of a choline-deficient diet during pregnancy and early postnatal life on the skeletal muscle transcriptome of the offspring.

BACKGROUND & AIMS: Nonalcoholic fatty liver disease (NAFLD) is related to muscle loss, but the precise mechanism underlying this association remains unclear. The aim of the present study was thus to determine the influence of maternal fatty liver and dietary choline deficiency during pregnancy and/or lactation periods on the skeletal muscle gene expression profile among 24-day-old male rat offspring. METHODS: Histological examination of skeletal muscle tissue specimens obtained from offspring of dams suffering from fatty liver, provided with proper choline intake during pregnancy and lactation (NN), fed a choline-deficient diet during both periods (DD), deprived of choline only during pregnancy (DN), or only during lactation (ND), was performed. The global transcriptome pattern was assessed using a microarray approach (Affymetrix® Rat Gene 2.1 ST Array Strip). The relative expression of selected genes was validated by real-time PCR (qPCR). RESULTS: Morphological differences in fat accumulation in skeletal muscle related to choline supply were observed. The global gene expression profile was consistent with abnormal morphological changes. Mettl21c gene was overexpressed in all choline-deficient groups compared to the NN group, while two genes, Cdkn1a and S100a4, were downregulated. Processes of protein biosynthesis were upregulated, and processes related to cell proliferation and lipid metabolism were inhibited in DD, DN, and ND groups compared to the NN group. CONCLUSIONS: Prenatal and early postnatal exposure to fatty liver and dietary choline deficiency leads to changes in the transcriptome profile in skeletal muscle of 24-day old male rat offspring and is associated with muscle damage, but the mechanism of it seems to be different at different developmental stages of life. Adequate choline intake during pregnancy and lactation can prevent severe muscle disturbance in the progeny of females suffering from fatty liver.

Foot-and-mouth disease-associated myocarditis is age dependent in suckling calves.

Myocarditis is considered a fatal form of foot-and-mouth disease (FMD) in suckling calves. In the present study, a total of 17 calves under 4 months of age and suspected clinically for FMD were examined for clinical lesions, respiratory rate, heart rate, and heart rhythm. Lesion samples, saliva, nasal swabs, and whole blood were collected from suspected calves and subjected to Sandwich ELISA and reverse transcription multiplex polymerase chain reaction (RT-mPCR) for detection and serotyping of FMD virus (FMDV). The samples were found to be positive for FMDV serotype "O". Myocarditis was suspected in 6 calves based on tachypnoea, tachycardia, and gallop rhythm. Serum aspartate aminotransferase (AST), creatinine kinase myocardial band (CK-MB) and lactate dehydrogenase (LDH), and cardiac troponins (cTnI) were measured. Mean serum AST, cTn-I and LDH were significantly higher (P < 0.001) in < 2 months old FMD-infected calves showing clinical signs suggestive of myocarditis (264.833 ± 4.16; 11.650 ± 0.34 and 1213.33 ± 29.06) than those without myocarditis (< 2 months old: 110.00 ± 0.00, 0.06 ± 0.00, 1050.00 ± 0.00; > 2 months < 4 months: 83.00 ± 3.00, 0.05 ± 0.02, 1159.00 ± 27.63) and healthy control groups (< 2 months old: 67.50 ± 3.10, 0.047 ± 0.01, 1120.00 ± 31.62; > 2 months < 4 months: 72.83 ± 2.09, 0.47 ± 0.00, 1160.00 ± 18.44). However, mean serum CK-MB did not differ significantly amongst the groups. Four calves under 2 months old died and a necropsy revealed the presence of a pathognomic gross lesion of the myocardial form of FMD known as "tigroid heart". Histopathology confirmed myocarditis. This study also reports the relevance of clinical and histopathological findings and biochemical markers in diagnosing FMD-related myocarditis in suckling calves.

Application of Statistical Analysis and Machine Learning to Identify Infants' Abnormal Suckling Behavior.

OBJECTIVE: Identify infants with abnormal suckling behavior from simple non-nutritive suckling devices. BACKGROUND: While it is well known breastfeeding is beneficial to the health of both mothers and infants, breastfeeding ceases in 75 percent of mother-child dyads by 6 months. The current standard of care lacks objective measurements to screen infant suckling abnormalities within the first few days of life, a critical time to establish milk supply and successful breastfeeding practices. MATERIALS AND METHODS: A non-nutritive suckling vacuum measurement system, previously developed by the authors, is used to gather data from 91 healthy full-term infants under thirty days old. Non-nutritive suckling was recorded for a duration of sixty seconds. We establish normative data for the mean suck vacuum, maximum suck vacuum, suckling frequency, burst duration, sucks per burst, and vacuum signal shape. We then apply computational methods (Mahalanobis distance, KNN) to detect anomalies in the data to identify infants with abnormal suckling. We finally provide case studies of healthy newborn infants and infants diagnosed with ankyloglossia. RESULTS: In a series of case evaluations, we demonstrate the ability to detect abnormal suckling behavior using statistical analysis and machine learning. We evaluate cases of ankyloglossia to determine how oral dysfunction and surgical interventions affect non-nutritive suckling measurements. CONCLUSIONS: Statistical analysis (Mahalanobis Distance) and machine learning [K nearest neighbor (KNN)] can be viable approaches to rapidly interpret infant suckling measurements. Particularly in practices using the digital suck assessment with a gloved finger, it can provide a more objective, early stage screening method to identify abnormal infant suckling vacuum. This approach for identifying those at risk for breastfeeding complications is crucial to complement complex emerging clinical evaluation technology. CLINICAL IMPACT: By analyzing non-nutritive suckling using computational methods, we demonstrate the ability to detect abnormal and normal behavior in infant suckling that can inform breastfeeding intervention pathways in clinic.Clinical and Translational Impact Statement: The work serves to shed light on the lack of consensus for determining appropriate intervention pathways for infant oral dysfunction. We demonstrate using statistical analysis and machine learning that normal and abnormal infant suckling can be identified and used in determining if surgical intervention is a necessary solution to resolve infant feeding difficulties.

A simple protocol to establish a conditionally immortalized mouse podocyte cell line.

Podocytes are specialized terminally differentiated cells in the glomerulus that are the primary target cells in many glomerular diseases. However, the current podocyte cell lines suffer from prolonged in vitro differentiation and limited survival time, which impede research progress. Therefore, it is necessary to establish a cell line that exhibits superior performance and characteristics. We propose a simple protocol to obtain an immortalized mouse podocyte cell (MPC) line from suckling mouse kidneys. Primary podocytes were cultured in vitro and infected with the SV40 tsA58 gene to obtain immortalized MPCs. The podocytes were characterized using Western blotting and quantitative real-time PCR. Podocyte injury was examined using the Cell Counting Kit-8 assay and flow cytometry. First, we successfully isolated an MPC line and identified 39 °C as the optimal differentiation temperature. Compared to undifferentiated MPCs, the expression of WT1 and synaptopodin was upregulated in differentiated MPCs. Second, the MPCs ceased proliferating at a nonpermissive temperature after day 4, and podocyte-specific proteins were expressed normally after at least 15 passages. Finally, podocyte injury models were induced to simulate podocyte injury in vitro. In summary, we provide a simple and popularized protocol to establish a conditionally immortalized MPC, which is a powerful tool for the study of podocytes.

Maternal immunization and vitamin A sufficiency impact sow primary adaptive immunity and passive protection to nursing piglets against porcine epidemic diarrhea virus infection.

Porcine epidemic diarrhea virus (PEDV) causes a highly contagious enteric disease with major economic losses to swine production worldwide. Due to the immaturity of the neonatal piglet immune system and given the high virulence of PEDV, improving passive lactogenic immunity is the best approach to protect suckling piglets against the lethal infection. We tested whether oral vitamin A (VA) supplementation and PEDV exposure of gestating and lactating VA-deficient (VAD) sows would enhance their primary immune responses and boost passive lactogenic protection against the PEDV challenge of their piglets. We demonstrated that PEDV inoculation of pregnant VAD sows in the third trimester provided higher levels of lactogenic protection of piglets as demonstrated by >87% survival rates of their litters compared with <10% in mock litters and that VA supplementation to VAD sows further improved the piglets' survival rates to >98%. We observed significantly elevated PEDV IgA and IgG antibody (Ab) titers and Ab-secreting cells (ASCs) in VA-sufficient (VAS)+PEDV and VAD+VA+PEDV sows, with the latter maintaining higher Ab titers in blood prior to parturition and in blood and milk throughout lactation. The litters of VAD+VA+PEDV sows also had the highest serum PEDV-neutralizing Ab titers at piglet post-challenge days (PCD) 0 and 7, coinciding with higher PEDV IgA ASCs and Ab titers in the blood and milk of their sows, suggesting an immunomodulatory role of VA in sows. Thus, sows that delivered sufficient lactogenic immunity to their piglets provided the highest passive protection against the PEDV challenge. Maternal immunization during pregnancy (± VA) and VA sufficiency enhanced the sow primary immune responses, expression of gut-mammary gland trafficking molecules, and passive protection of their offspring. Our findings are relevant to understanding the role of VA in the Ab responses to oral attenuated vaccines that are critical for successful maternal vaccination programs against enteric infections in infants and young animals.

Maternal behavior of dairy cows and suckling behavior of dairy calves in different cow-calf contact conditions.

Cow-calf contact has been suggested as an alternative to the common practice of early separation of dairy cow and calf. Increasing cow-calf contact poses challenges by reducing the machine milk yield and increasing separation stress. Part-time cow-calf contact may prove more feasible than full-time contact for farmers but may affect the maternal behavior of the cows. Gradually reducing the daily contact could gradually wean the calf off milk, preparing it for separation. This study aims to investigate (1) how cows' maternal behaviors toward their own calf are affected by part-time contact, and (2) whether calves' suckling behavior is reduced by reducing daily cow contact. Fifty-six dairy cows and their calves were assigned to either full-time (23 h/d) or part-time (10 h/d) cow-calf contact for 7 wk (main rearing period). This was followed by 2 wk of either reduced contact (50% in wk 8, then 25% of original contact time in wk 9) or unchanged contact. First, the maternal behavior of full- and part-time cows was observed for 24 h in the third week of the main rearing period. Part-time cows spent less time nursing and grooming their own calf than full-time cows did. However, the amount of nursing in the inverse parallel position and the probability of a cow nursing a calf other than her own did not differ between the 2 treatments. Second, calf suckling behavior was observed for 24 h in both weeks of the reduced contact period. Averaged across both observations, total suckling time (i.e., time spent suckling any cow) was lower in part-time calves that experienced reduced contact compared with part-time calves with unchanged contact. In wk 9, time spent suckling only the dam was lower for full-time calves with reduced contact compared with part-time calves with unchanged contact. In conclusion, although we found a lower nursing and grooming time in part-time cows compared with full-time, we found no difference between treatments with regard to nursing in the inverse parallel position and nursing a calf other than the cow's own. The effect of gradually reducing cow-calf contact on calves' suckling time requires more research.

Bacillus subtilis alleviates excessive apoptosis of intestinal epithelial cells in intrauterine growth restriction suckling piglets via the members of Bcl-2 and caspase families.

BACKGROUND: The intestine is a barrier resisting various stress responses. Intrauterine growth restriction (IUGR) can cause damage to the intestinal barrier via destroying the balance of intestinal epithelial cells' proliferation and apoptosis. Bacillus subtilis has been reported to regulate intestinal epithelial cells' proliferation and apoptosis. Thus, the purpose of this study was to determine if B. subtilis could regulate intestinal epithelial cells' proliferation and apoptosis in intrauterine growth restriction suckling piglets. RESULTS: Compared with the normal birth weight group, the IUGR group showed greater mean optical density values of Ki-67-positive cells in the ileal crypt (P < 0.05). IUGR resulted in higher ability of proliferation and apoptosis of intestinal epithelial cells, by upregulation of the messenger RNA (mRNA) or proteins expression of leucine rich repeat containing G protein coupled receptor 5, Caspase-3, Caspase-7, ß-catenin, cyclinD1, B-cell lymphoma-2 associated agonist of cell death, and BCL2 associated X (P < 0.05), and downregulation of the mRNA or protein expression of B-cell lymphoma-2 and B-cell lymphoma-2-like 1 (P < 0.05). However, B. subtilis supplementation decreased the mRNA or proteins expression of leucine rich repeat containing G protein coupled receptor 5, SPARC related modular calcium binding 2, tumor necrosis factor receptor superfamily member 19, cyclinD1, Caspase-7, ß-catenin, B-cell lymphoma-2 associated agonist of cell death, and Caspase-3 (P < 0.05), and increased the mRNA expression of B-cell lymphoma-2 (P < 0.05). CONCLUSION: IUGR led to excessive apoptosis of intestinal epithelial cells, which induced compensatory proliferation. However, B. subtilis treatment prevented intestinal epithelial cells of IUGR suckling piglets from excessive apoptosis. © 2024 Society of Chemical Industry.

Effective breastfeeding techniques and associated factors among lactating women: a community-based study, north east Ethiopia.

Background: Effective breastfeeding techniques, which include proper attachment, positioning, and suckling, offer a range of benefits for both the mother and the infant. These techniques ensure efficient milk transfer, reduce the risk of infections, support optimal infant weight gain, enhance maternal comfort, and foster a strong emotional bond. This study aimed to identify the magnitude and factors associated with effective breastfeeding techniques among lactating women in the Legambo district of South Wollo, Ethiopia, in 2022. Methods: A community-based cross-sectional study was conducted from September to November 2022. Samples were selected using a multi-stage sampling method from 18 wards (kebele). Data were collected using an interviewer-administered structured questionnaire and an observational checklist. The collected data were entered into Epi-Data and then exported to SPSS version 25.0 for analysis. Descriptive statistics and bivariate and multivariable logistic regression analyses were performed to identify the magnitude and associated factors. Variables with a p-value less than 0.05 on multivariable analysis were considered independent factors associated with the outcome variable. Results: Six hundred and ten lactating women were included for observation and interviewed, resulting in a 96.2% response rate. The magnitude of effective breastfeeding technique practice was found to be 25.9% (95% CI: 22.47-29.57%). Factors associated with effective breastfeeding technique practice included being a working woman (AOR = 1.70; 95%CI: 1.07-2.72), age between 26 and 30 years (AOR = 0.37; 95%CI: 0.16-0.84), urban residence (AOR = 1.59; 95%CI: 1.06-2.39), initiating breastfeeding 1 to 2 h after birth (AOR = 0.27; 95%CI: 0.16-0.43), and initiating breastfeeding after 2 h of birth (AOR = 0.34; 95%CI: 0.17-0.67). Additionally, not receiving breastfeeding education (AOR = 0.46; 95%CI: 0.30-0.72) and experiencing current breast problems (AOR = 0.28; 95%CI: 0.28-0.75) were also found to have a significant association with effective breastfeeding technique practice. Conclusion: Only one in four women demonstrated effective breastfeeding techniques, indicating that their practice was below the WHO's recommendations. Therefore, it is crucial to consider the identified variables to improve the practice of effective breastfeeding techniques.

Observation of the Effects of Different Enzyme Digestion Time and Enzyme Digestion Methods on the Extraction of Primary Renal Tubular Epithelial Cells from Sprague-Dawley Suckling Mice.

Renal tubular epithelial cells are one of the essential functional cells in the kidney. Optimizing the isolation and culture method of primary renal tubular epithelial cells from SD mammary rats provides better experimental materials for renal tubule-related studies, which is essential for studying the pathogenesis of renal diseases, especially diabetic nephropathy and drug screening. SD rat renal tubular epithelial cells were isolated and purified by 2.5-mg/ml collagenase II or 2 mg/ml trypsin + 2.5 mg/ml collagenase II enzymatic digestion. The isolation and purification were observed at different time points (15 min, 30 min, 45 min, and 60 min) to determine the optimal extraction time for the enzymatic digestion method. After comparing the two enzymatic methods, it was determined that the trypsin + collagenase II enzymatic method was more effective. The primary renal tubular epithelial cells extracted by the trypsin + collagenase II digestion method were identified by the marker Cytokeratin 18 of renal tubular epithelial cells at 45 min of digestion with high purity. We established a simple, efficient, and reproducible method for isolation and culture of renal tubular epithelial cells in SD mammary gland rats.