Anaerobic flora, Selenomonas ruminis sp. nov., and the bacteriocinogenic Ligilactobacillus salivarius strain MP3 from crossbred-lactating goats.

This study aimed to examine the distribution of anaerobic bacteria in the rumen fluid of Thai crossbred goats and to screen potential probiotic strains capable of producing antimicrobial compounds and inhibiting bacteria that cause milk fat depression. Thirty-four strains of bacteria from the rumen fluid were divided into 13 groups within 12 genera based on 16S rRNA gene sequences. The RF1-5 and RF5-12 were identified as Streptococcus luteliensis and Bacillus licheniformis, respectively, and demonstrated non-ropy exopolysaccharide. Furthermore, mPRGC5T was closely related to Selenomonas caprae JCM 33725 T (97.8% similarity) based on 16S rRNA gene sequences. It exhibited low average nucleotide identity, digital DNA-DNA hybridization, and average amino acid identity values with related type strains ranging from 84.9 to 86.0%, 21.3 to 21.8%, and 73.8 to 76.1%, respectively. The genotypic and phenotypic characteristics of mPRGC5T strongly support this strain as a new species of the genus Selenomonas for which the name Selenomonas ruminis mPRGC5T was proposed. The type strain is mPRGC5T (= JCM 33724 T = KCTC 25177 T). Ligilactobacillus salivarius MP3 showed antibacterial activity against Cutibacterium acnes subsp. acnes DSM 1897 T and Kocuria rhizophila MIII. The enterolysin A cluster gene was identified in its genome. The auto-aggregation of L. salivarius MP3 was 93.6 ± 0.2%. Additionally, co-aggregation of L. salivarius MP3 with C. acnes DSM 1897 T and K. rhizophila MIII had 92.2 ± 3.4% and 87.3 ± 4.5%, respectively. The adhesion capacity of strain MP3 was 76.11 ± 2.2%. Probiogenomic analysis revealed that L. salivarius MP3 was nonhazardous to animal supplementation and included acid- and bile-tolerant ability. However, strain MP3 contained three antibiotic resistance genes. Thus, the supplementation of L. salivarius MP3 could increase the milk fat content by suppressing C. acnes DSM 1897 T with antibiotic resistance gene horizontal transfer awareness.

Selenomonas caprae sp. nov., an obligately anaerobic and volatile fatty acid producing bacterium from ruminal fluid of domestic goat (Capra hircus L.).

A novel Gram-stain-negative, motile, obligately anaerobic bacterium strain mPRGC8T was isolated from the ruminal fluid of a domestic goat (Capra hircus L.) in Nakhon Pathom province, Thailand. The strain grew at 20-45 °C (optimum, 37 °C), pH 6.0-9.0 (optimum, pH 7.5) and 3 % (w/v) NaCl. It produced acetate, propionate, valerate, caproate and heptanoate from glucose. The 16S rRNA gene sequence analysis indicated that strain mPRGC8T belonged to the genus Selenomonas and was closely related to Selenomonas ruminantium subsp. ruminantium DSM 2150T (98.0 %) and Selenomonas ruminantium subsp. lactilytica JCM 6582T (97.9 %). The in silico DNA G+C content was 53.0 mol %. Strain mPRGC8T showed average nucleotide identity, digital DNA-DNA hybridization and average animo acid identity values with Selenomonas montiformis JCM 34373T, S. ruminantium subsp. lactilytica JCM 6582T and S. ruminantium subsp. ruminantium DSM 2150T ranging from 84.9 to 86.0 %, 21.3 to 21.8 % and 73.8 to 76.1 %, respectively. The predominant cellular fatty acids were C16 : 1 ω9c and C18 : 1 ω9c. Phosphatidylethanolamine, three unidentified aminophospholipids, two unidentified ninhydrin positive glycolipids, an unidentified phospholipid and an unidentified lipid were detected as polar lipids. The genomic and phenotypic characteristics of strain mPRGC8T strongly support its classification as representative of new species of the genus Selenomonas for which the name Selenomonas caprae sp. nov. is proposed. The type strain is mPRGC8T (=JCM 33725T=KCTC 25178T).

High dietary cation and anion difference formulation increased heat dissipation in non-lactating goats fed at high ambient temperature.

Background and Aim: In our previous study, we observed that a high dietary cation and anion difference (DCAD) of 40 mEq/kg dry matter (DM) in the diets of lactating dairy goats increased heat dissipation. In the present study, we believe that the level of DCAD fed to non-lactating and non-pregnant goats was twice as high as that fed to lactating goats in our previous study. This increase could have resulted in a greater water balance due to increased intake of water and unchanged urinary excretion. Therefore, this study aimed to determine the behavioral and heat dissipation effects of a dietary shift from low to high DCAD levels in dairy goats under tropical conditions. Materials and Methods: Seven non-lactating and non-pregnant crossbred goats were used in this study. All animals were initially fed a low DCAD (15 mEq/100 g DM) diet from days 0-6 and then switched to a high DCAD (89 mEq/100 g DM) diet from day 7 (high DCAD-7) to day 18 (high DCAD-18). Results: The results revealed that a high DCAD increased DM intake from days 13-18 (p < 0.05). The larger daily meal size associated with the high DCAD-18 group was due to increased daytime meal sizes, not nighttime when compared to the low DCAD group. Dietary cation and anion difference supplementation did not affect daily water intake; however, drinking patterns differed between the low DCAD group and the high DCAD-7 group from 07:00 to 09:00 and during nighttime. Similarly, daily urine volume was unaffected by DCAD supplementation, but urinary patterns differed between the low DCAD and high DCAD-18 groups. The daily water balance remained unchanged across all treatments, yet, a higher morning water balance was observed in the high DCAD group. The high DCAD diet led to an increase in respiration rate and rectal temperature compared to the low DCAD diet. Conclusion: The observed eating, drinking, and urinary patterns collectively suggested that high DCAD supplementation mitigates the effects of heat stress in non-lactating goats fed at high ambient temperatures.

Seasonal effect of milk yield and blood metabolites in relation to ketosis of dairy cows fed under a high ambient temperature.

BACKGROUND AND AIM: Metabolism and environment are closely related. Under high ambient temperature (HTa), dairy cows may have different energy metabolism during summer and winter. The present study was carried out to investigate the effect of HTa on the milk yield and blood concentration of beta-hydroxybutyrate (BHBA) and glucose at the herd level. MATERIALS AND METHODS: One large dairy farm in Thailand with more than 100 crossbred Holstein cows milked each month was selected. The first experiment was performed on non-lactating cows to determine the normal daily concentrations of blood BHBA and glucose. Under the HTa condition, there was no significant change in blood BHBA and glucose concentrations. The second experiment was performed using a prospective cohort clinical design to demonstrate the seasonal effect on milk yield and blood BHBA as an indication of energy metabolism at the herd level. RESULTS: The temperature and humidity index for the winter (78.1±0.5) and summer (83.4±0.7) periods differ significantly. The average milk yield during the winter period was 17.8% higher than during the summer period. The reduction of body condition score (BCS) during early lactation was significant in the winter cows. Both higher milk yield and lower BCS in the winter cows suggested a state of negative energy balance. However, there was no difference in blood BHBA and glucose concentrations between winter and summer cows. The effect of HTa on insulin signaling appeared to be a counterbalancing factor for the ketogenic status. Based on the present results, it would be interesting to further investigate the incidence of subclinical and clinical ketosis in a dairy farm under tropical conditions. CONCLUSION: The present experiment revealed that HTa during summer decreased milk yield in dairy cows fed under tropical conditions. Higher milk yield in winter caused a greater reduction of BCS and suggested a greater negative energy balance. However, there was no seasonal effect on blood BHBA and glucose concentrations.

Exogenous bovine somatotropin and mist-fan cooling synergistically promote the intramammary glucose transport for lactose synthesis in crossbred Holstein cows in the tropics.

BACKGROUND AND AIM: Milk synthesis by the mammary gland is negatively influenced in part by high ambient temperature (AT). This study aimed to clarify the pathway of intramammary glucose utilization involved in mediating lactose synthesis during treatment with somatotropin under housing with misters and fans. MATERIALS AND METHODS: A single subcutaneous injection of 500 mg of recombinant bovine somatotropin (rbST) was administered 3 times once every 14 days to 87.5% crossbred Holstein cattle in early-/mid-/late lactation, under housing in a normal shaded barn and in a shaded barn with a mist-fan cooling system. RESULTS: The milk yields of the cooled cows tended to increase compared with those of uncooled cows and exhibited more potentiated effects in response to rbST treatment, coinciding with increases in mammary plasma flow and glucose uptake, but not in the mammary extraction of glucose. Treatment with rbST in the cooled cows resulted in a greater increase in the milk glucose concentration and a greater decrease in the milk glucose-6-phosphate concentration at all stages of lactation. CONCLUSION: rbST treatment exerted its galactopoietic action more by local intramammary factors than by other extramammary factors at a low AT and the synergistic effect between rbST treatment and low AT increased the availability of intramammary glucose transport in activating the process of lactose synthesis.

Effects of dietary cation and anion difference on eating, ruminal function and plasma leptin in goats under tropical condition.

OBJECTIVE: This study was carried out to determine the effects of elevated dietary cation and anion difference (DCAD) on dry matter intake (DMI) and ruminal fermentation pattern in lactating dairy goats under tropical conditions. METHODS: Ten dairy goats were divided into two groups of five animals each. The groups received diets at different DCAD levels, either a control diet (22.81 mEq/100 g dry matter [DM], DCAD-23) or a DCAD-39 diet (39.08 mEq/100 g DM, DCAD-39). After parturition, DMI and water intake were recorded daily. Ruminal fluid and urine were collected, and nutrient digestibility measurements were carried out at 8th weeks postpartum (PP-8). Blood samples were collected at PP-4 and PP-8 to measure plasma leptin. RESULTS: Dry matter intake/body weight (DMI/BW) at PP-8 of the animals fed the DCAD-39 diet was significantly higher than those fed with DCAD-23 diet (p<0.05). Animals fed with DCAD-39 consumed more water than those fed DCAD-23 over 24 h, particularly at night (p<0.05). Ruminal pH, acetate concentration, and urinary allantoin excretion increased with the DCAD-39 diet, whereas ruminal butyrate concentration was lower with the DCAD-39 diet. On the other hand, other ruminal parameters, such as total volatile fatty acid concentration, propionate molar proportion and acetate/propionate average ratio, were not affected by increased DCAD supplementation. Apparent digestibility was improved by increased DCAD supplementation. Plasma leptin concentration was higher with DCAD supplementation. CONCLUSION: When feeding goats with DCAD-39 under tropical conditions, an increase in DMI was associated with improved apparent digestibility of nutrients, ruminal fermentation and microbial protein synthesis. An increase in plasma leptin concentration could not explain the effect of high DCAD on DMI.

The effect of dietary ions difference on drinking and eating patterns in dairy goats under high ambient temperature.

OBJECTIVE: The present study was carried out to evaluate the effect of high dietary cation and anion difference (DCAD) rations on diurnal variations in eating and meal patterns, water intake and urination patterns in dairy goats fed under high ambient temperature (HTa). METHODS: Ten crossbred dairy goats during peri-parturition period were selected and divided into two groups of five animals each. Experimental diets were control DCAD (control, 22.8 mEq/100 g dry matter [DM]) and high DCAD (DCAD, 39.1 mEq/100 g DM). The composition of two diets consisted of 44% corn silage and 56% concentrate. From the 2nd week to 8th week postpartum, goats were fed ad libitum twice daily either with the control or DCAD total mix ration with free access to water. The spontaneous eating and drinking patterns were determined. RESULTS: The environmental conditions in the present experiment indicated that goats were fed under HTa conditions (average peak THI = 85.2) and were in heat stress. In addition to the typical HTa induced tachypnoea in both groups, the respiratory rate in the DCAD group was significantly higher than the control group (p<0.05). Although the goats from both groups showed comparable level of eating, drinking and urination during experiment, the meal pattern and water intake were different. High DCAD apparently increased eating and meal patterns compared with the control. At week 8 postpartum, goats from high DCAD group had significant (p<0.05) bigger meal size and longer meal duration. Moreover, high DCAD appeared to increase night-time water intake (p<0.05). CONCLUSION: Both meal pattern and night-time drinking effects of DCAD suggested that feeding with high DCAD ration may alleviate the effect of heat stress in dairy goat fed under HTa conditions.

Dietary cation and anion difference: Effects on milk production and body fluid distribution in lactating dairy goats under tropical conditions.

This study aimed to determine the effect of dietary cation and anion difference (DCAD) on milk production and body fluid distribution in lactating dairy goats. Ten dairy goats were selected and divided into two groups, five animals each. Animals received either control DCAD (control, 22.81 mEq/100 g dry matter (DM)) or high DCAD (DCAD, 39.08 mEq/100 g DM). The results indicated that rectal temperature (Tr), respiration rate, milk yield and compositions did not differ between groups. But the percentage change of Tr from the DCAD group was lower than the control group between 09.00 and 13.00 hours. DM intake tended to increase in the DCAD group. Dairy goats in the DCAD group drank more water, but urinary excretion and plasma antidiuretic hormone concentration remained unchanged. Apparent water balance was higher from the DCAD group over 24 h. There was no effect of DCAD on plasma and blood volumes, but tended to increase in extracellular fluid and thereby increased total body water. The present results indicate that animals supplemented with high DCAD increase their total body water and apparent water balance. These results have contributed to the process of adaptation for evaporative cooling and would be useful in slowing down the elevation in Tr.

The association of ruminal pH and some metabolic parameters with conception rate at first artificial insemination in Thai dairy cows.

The objective of this study was to determine the association of metabolic parameters and cow associated factors with the conception rate at first insemination (FCR) in Thai dairy cows. The investigation was performed with 529 lactations from 32 smallholder dairy farms. At 3-6 weeks after parturition, blood samples and ruminal fluid were collected. Body condition scores (BCS) of cows were scored 1 week before expected calving date and at blood sampling date. Ruminal pH was measured at 2-4 h after morning feeding in ruminal fluid collected by ruminocentesis. Serum beta-hydroxybutyrate and serum urea nitrogen were measured by kinetic enzyme method. Cows with first insemination (AI) between 41 and 114 days postpartum were identified after pregnancy diagnosis for FCR. Breed, parity, interval from calving to first AI, BCS before calving, BCS after calving, loss in BCS after calving, SBHB, SUN, ruminal pH, and postpartum problems were selected as independent variables for a model with FCR as a dependent variable. A multivariable logistic regression model was used with farm as a random effect. Overall FCR was 27.2 %. The FCR depended on interval from calving to first AI, BCS before calving, and ruminal pH. The FCR between 69 and 91 days postpartum was significantly highest (45 %). Before calving, a cow with high BCS (≥ 3.5) had significantly greater FCR than a cow with low BCS (≤ 3.25; P<0.01). An increased ruminal pH raised significantly FCR (OR=2.53; P=0.03).

Effects of supplemental recombinant bovine somatotropin and mist-fan cooling on the renal tubular handling of sodium in different stages of lactation in crossbred Holstein cattle.

The effect of supplementary administration of recombinant bovine somatotrophin (rbST) on the renal tubular handling of sodium in crossbred 87.5% Holstein cattle housed in normal shade (NS) or mist-fan cooled (MF) barns was evaluated. The cows were injected with 500 mg rbST at three different stages of lactation. The MF barn housed cows showed a slightly decreased ambient temperature and temperature humidity index, but an increased relative humidity. Rectal temperature and respiration rates were significantly lower in cooled cows. The rbST treated cows, housed in NS or MF barns, showed markedly increased milk yields, total body water, extracellular fluid and plasma volume levels, along with a reduced rate of urine flow and urinary excretion of sodium, potassium and chloride ions and osmolar clearance, in all three stages of lactation. Renal tubular sodium and water reabsorption were increased after rbST administration without any alteration in the renal hemodynamics. Lithium clearance data suggested that the site of response is in the proximal nephron segment, which may be mediated via increases in the plasma levels of aldosterone and IGF-1, but not vasopressin, during rbST administration.

Effects of misty-fan cooling and supplemental rbST on rumen function and milk production of crossbred Holstein cattle during early, mid and late lactation in a tropical environment.

Two groups of five crossbred 87.5% Holstein cattle were housed in normal shade only (NS) as non-cooled cows and in shaded housing with misty-fan cooling (MF) as cooled cows. The cows were treated with recombinant bovine somatotropin (rbST) in early, mid and late lactation with three consecutive injections of rbST in every 14 days. Ambient temperatures and the temperature humidity index in the NS barn were significantly higher than those of the MF barn, whereas relative humidity in MF was higher than that of NS barn. The DMI of cooled cows were higher than those of non-cooled cows, and cooled cows exhibited more response to rbST treatment. Exogenous rbST significantly increased milk yield throughout lactation. The rbST-treated cows had higher total ruminal fermentation products as volatile fatty acid and ammonia nitrogen than the non-rbST treated cows and associated changes were greater in cooled animals in all stages of lactation. Exogenous rbST increased the concentrations of milk urea nitrogen in both groups. These results suggest that the changes in ruminal fermentation with greater production of total VFA and NH(3)N in response to rbST in crossbred cows whether under misty-fan cooling or not, is in part through an increase in feed intake, thereby making more substrate available to the mammary gland for milk synthesis.