Matrices-charges of agar-alginate crosslinked microcapsules via o/w microemulsion: A non-spore forming probiotic bacteria encapsulation system for extensive viability.

Non-spore forming probiotic bacteria tend to diminish their activity under adverse conditions. This leads to the difficulty of delivery in animal body system as well as the feed pelleting process. The present work proposes the microcapsule networks based on polymer matrices and charges under ionic crosslink to encapsulate probiotic for an extensive stability in adverse conditions. The combination of agar (AG) and alginate (AL) is a good model to combine agar matrices and alginate charges under ionic crosslink through o/w emulsion system for probiotic incorporation. By simply mixing Lactobacillus plantarum MB001(LPMB001) with AL and AG containing few drops of soybean oil, a stable o/w microemulsion can be obtained. The addition of calcium chloride favors the ionic crosslink among AG matrices resulting in LPMB001/AG-AL microcapsules. In vitro studies indicate the survival of LPMB001 and the slow release even after treatment in adverse conditions. This microencapsulation prolongs LPMB001 viability under the heat treatment and the storage conditions and this designates the potential feed processing. The present work for the first time shows how we can combine polymer matrices and charges to protect probiotic from the adverse conditions which is simple and practical for the process of dietary supplementation.

Microencapsulation of Lactobacillus plantarum MB001 and its probiotic effect on growth performance, cecal microbiome and gut integrity of broiler chickens in a tropical climate.

OBJECTIVE: Microencapsulation technologies have been developed and successfully applied to protect the probiotic bacterial cells damaged by environmental exposure. This study aimed to investigate the effects of microencapsulation of Lactobacillus plantarum MB001 on the growth performance, ileal nutrient digestibility, jejunal histomorphology and cecal microbiome of broiler chickens in a tropical climate. METHODS: A total of 288 one-day-old female broilers (Ross 308) were randomly allocated into 4 groups (6 replicates of 12 birds). Treatments included, i) a basal diet (NC), ii) NC + avilamycin (10 mg/kg) (PC), iii) NC + non-encapsulated L. plantarum MB001 (1×108 colonyforming unit [CFU]/kg of diet) (N-LP), iv) NC + microencapsulated L. plantarum MB001 (1×108 CFU/kg of diet) (ME-LP). RESULTS: Dietary supplementation of ME-LP improved average daily gain, and feed conversion ratio of broilers throughout the 42-d trial period (p<0.05), whereas ME-LP did not affect average daily feed intake compared with NC group. Both N-LP and ME-LP improved apparent ileal digestibility of crude protein and ether extract compared with NC group (p<0.05). The broilers fed ME-LP supplemented diet exhibited a beneficial effect on jejunal histomorphology of villus height (VH), crypt depth (CD) and villus height to crypt depth ratio (VH:CD) of broilers compared to NC group (p<0.05). At the phylum level, Firmicutes was enriched (p<0.05) and Proteobacteria was decreased (p<0.05) only in the ME-LP group. At the genus level, the ME-LP diets increased (p<0.05) the number of both Lactobacillus and Enterococcus compared to NC, PC, and N-LP groups (p<0.05). CONCLUSION: Microencapsulation assists the efficient functioning of probiotics. ME-LP could be potentially used as a feed additive for improvement of cecal microbiota, gut integrity and nutrient utilization, leading to better performance of broilers.

Retrospective analysis of canine monocytic ehrlichiosis in Thailand with emphasis on hematological and ultrasonographic changes.

Background and Aim: Canine monocytic ehrlichiosis (CME) is a tropical endemic tick-borne disease that causes fatality or chronic infection involving many organs in dogs. This study aimed to examine the prevalence, risk factors, and hematological and ultrasonographic changes in the liver, gallbladder, kidneys, and spleen following CME infection. Materials and Methods: This retrospective study used 30,269 samples collected from dogs at the hematology section of the pathology unit of a university veterinary hospital and 35 samples collected from dogs at the diagnostic imaging unit. CME was determined using the buffy coat smear method. Data were analyzed using descriptive statistics and odds ratios. Results: The data revealed that the average yearly prevalence of CME was 1.32%. Risk factors contributing to CME infection were a tick on the body during physical examination, lack of ectoparasite control, and outdoor living. All 148 dogs with CME infection had low platelet counts. The percentages of CME-infected dogs with elevated serum alanine aminotransferase, alkaline phosphatase, and both enzymes above the normal range were 33.6%, 65.9%, and 29.8%, respectively. The rates for elevated serum levels of blood urea nitrogen, creatinine, and both compounds were 33.1%, 19.1%, and 17.3%, respectively. The most common ultrasonographic changes were liver abnormalities (hyperechogenicity or hypoechogenicity, hepatomegaly, and hypoechoic nodules), hyperechogenicity of the kidneys, and an enlarged spleen. These ultrasonographic changes were consistent with the hematology results, which showed a greater elevation of serum liver enzyme levels than renal enzymes. Conclusion: Ultrasonographic changes during CME infection and after treatment with doxycycline can help to monitor and identify persistent pathological changes in the target organs resulting from immune response to CME.

Microencapsulated basil oil (Ocimum basilicum Linn.) enhances growth performance, intestinal morphology, and antioxidant capacity of broiler chickens in the tropics.

OBJECTIVE: Microencapsulation is a technique to improve stability, bioavailability, and controlled release of active ingredients at a target site. This experiment aimed to investigate the effects of microencapsulated basil oil (MBO) on growth performance, apparent ileal digestibility (AID), jejunal histomorphology, bacterial population as well as antioxidant capacity of broiler chickens in a tropical climate. METHODS: A total of 288 one-day-old female broilers (Ross 308) were randomly allocated into 4 groups (6 replicates of 12 birds), based on a completely randomized design. Dietary treatments were as follows: i) basal diet (NC), ii) basal diet with avilamycin at 10 ppm (PC), iii) basal diet with free basil oil (FBO) at 500 ppm, and iv) basal diet with MBO at 500 ppm, respectively. RESULTS: Dietary supplementation of MBO improved average daily gain, and feed conversion ratio of broilers throughout the 42-d trial period (p<0.05), whereas MBO did not affect average daily feed intake compared with NC group. The broilers fed MBO diet exhibited a greater AID of crude protein and gross energy compared with those in other groups (p<0.05). Lactobacillus spp. and Escherichia coli populations were not affected by feeding dietary treatments. Both FBO and MBO had positive effects on jejunal villus height (VH), villus height to crypt depth ratio (VH:CD) and villus surface area of broilers compared to NC and PC groups (p<0.05). Superoxide dismutase level in the duodenal mucosa of MBO group was significantly increased (p<0.01), whereas malondialdehyde level was significantly decreased (p<0.01). CONCLUSION: Microencapsulation could be considered as a promising driver of the basil oil efficiency, consequently MBO at 500 ppm could be potentially used as a feed additive for improvement of intestinal integrity and nutrient utilization, leading to better performance of broiler chickens.

A urinary proteomic study in hypercalciuric dogs with and without calcium oxalate urolithiasis.

Background and Aims: Hypercalciuria is an important predisposing factor commonly found in humans and dogs with calcium oxalate (CaOx) urolithiasis. Calcium oxalate crystals can induce an inflammatory reaction that subsequently produces several proteins that have an inhibitory or stimulatory effect on stone formation. This study aimed to evaluate the differences in urinary proteomic profiles between hypercalciuric CaOx stone dogs and hypercalciuric stone-free dogs (CaOx stone and control groups, respectively). Materials and Methods: Seven dogs with hypercalciuric CaOx urolithiasis and breed-, sex-, and aged-matched controls with hypercalciuria were included in the study. Serum and urine samples were obtained from all dogs to analyze electrolytes. Urinary proteomic profiles were analyzed using liquid chromatography-mass spectrometry. Student's t-test was used to compare the differences between groups. Results: Forty-nine urinary proteins were identified in the stone-free and CaOx stone groups, whereas 19 and 6 proteins were unique in the CaOx stone and stone-free groups, respectively. The urinary thrombomodulin level was significantly higher in the CaOx stone group (relative ratio = 1.8, p < 0.01) than in the stone-free group. Conclusion: This study demonstrated that urinary proteomic profiles may be used as a candidate biomarker for urinary tract injury in CaOx urolithiasis in dogs.

Comparison of fine-needle cytologic diagnosis between the left and right liver lobes of dogs and cats with diffuse liver disease.

BACKGROUND AND AIM: Ultrasound-guided fine-needle sample collection for cytology with manual restraint is frequently used for the primary assessment of diffuse liver disease in veterinary patients in Thailand. For better diagnosis, repeated collection of samples ensures the collection of adequate, representative samples, which increase diagnostic accuracy. However, in those that are unable to receive general anesthesia, it is difficult to collect the samples from several liver locations in manually restrained dogs and cats. The study aimed to compare the cytologic diagnosis of the ultrasound-guided fine-needle non-aspiration technique between the left and right liver lobes in dogs and cats with neoplastic and non-neoplastic diffuse liver disease. MATERIALS AND METHODS: This prospective study included 25 client-owned dogs and cats with diffuse liver diseases. Two liver samples were randomly collected from the left and right liver lobes under ultrasound guidance for cytologic examination. All slides were subsequently examined blindly by experienced pathologists for cytologic analysis with cytologic agreement scores (CASs). RESULTS: Among all 50 samples obtained from ultrasound-guided fine-needle sample collection of the left and right liver, 78% were diagnostic and 22% were non-diagnostic. In the diagnostic group, 73.3% of fine-needle samples had concordant results between the left and right liver, which exhibited 100% cytologic agreement in lymphoma and 63.6% in non-neoplastic groups. Samples collected from the left liver had slightly higher CAS and higher cytologic quality than had those from the right liver lobe (p=0.053). CONCLUSION: The location and number of sample collections did not have a significant difference in the cytologic diagnosis of diffuse liver disease, especially in patients with lymphoma. For manually restrained patients, one time ultrasound-guided non-aspiration cytology procedure from the left liver lobe not only decreased restraint duration and minimized tissue trauma but also allowed for an adequate cytologic diagnosis in diffuse liver disease compared to multiple collections.

The association between single nucleotide polymorphism in vitamin D receptor and calcium oxalate urolithiasis in dogs.

BACKGROUND: Polymorphisms of the vitamin D receptor (VDR) are associated with calcium oxalate (CaOx) nephrolithiasis in humans. OBJECTIVES: To investigate the association between VDR polymorphisms and susceptibility to CaOx urolithiasis in dogs. ANIMALS: Thirty-five dogs with CaOx urolithiasis were compared with 40 stone-free dogs. METHODS: This was a case-control study. Two VDR gene polymorphisms (rs851998024 and rs852900542) were detected by specific TaqMan real-time polymerase chain reaction assay, and their relationship with serum 1,25-dihydroxyvitamin D, serum and urinary electrolyte concentrations was evaluated. RESULTS: The distribution of the rs852900542 polymorphism was significantly different between the case and the control dogs (x2  = 6.369, P = .04). Dogs with a CC or CT genotype had an increased risk of CaOx stones than those with the TT genotype (odds ratio = 3.82, 95% confidence interval 1.04-13.98). The CaOx dogs with the TT genotype had a significantly lower urinary calcium-to-creatinine ratio than the CT+CC genotypes. 1,25-(OH)2D concentrations did not differ between the cases and the controls (308.7 ± 217.4 vs 286.7 ± 185.1 pg/mL, P = .45). CONCLUSIONS AND CLINICAL IMPORTANCE: This finding suggests that vitamin D metabolism might play a role in CaOx stone formation in dogs.

Effect of Dietary Inclusion of Gynura divaricata (L.) on Growth Performance, Hematology, and Carcass Fat Deposition in Broilers.

The objective of this study was to examine the effect of Gynura divaricata (Jakr-Na-Rai, JNR) on the growth performance, hematology, and carcass fat deposition of broilers. A total of 240 male Cobb-500 birds, 22 d old, were randomly allocated into five treatment groups of six replicates. Each group was raised at a high stocking density of 28 kg of bodyweight/m2 until day 43. The treatments consisted of (i) a basal diet, or the basal diet supplemented with (ii) 2.5 mg/kg avilamycin (T2), (iii) JNR crude ethanol extract at a flavonoid level of 1.3 g/kg (T3), and (iv and v) JNR powder at a flavonoid level of 1.3 (T4) or 2.6 g/kg (T5). Dietary supplementation with JNR powder at both flavonoid levels decreased the heterophil/lymphocyte ratio compared to the other groups (P<0.001). Birds in the T5 group presented decreased blood glucose (P<0.005) and cholesterol (P<0.002) levels and a numerically (P=0.056) decreased triglyceride level. The total bile acid concentration increased (P<0.001) in all the JNR-fed groups, but there was no significant effect on the digestibility of ileal protein or fat. At 1.3 g/kg of diet, JNR increased the final bodyweight and feed intake (P<0.05), but the average daily gain and feed conversion ratio were not different among groups. The carcass and abdominal fat percentages were lowest in chicks fed on the diets supplemented with 2.6 g/kg JNR powder (P<0.007 and P<0.025, respectively). Drip loss and malondialdehyde concentrations in the breast meat did not change. In conclusion, JNR powder improved several hematological parameters, increased total bile acid concentrations, and decreased the percentage of abdominal fat. The powder form of JNR elicited better results than the ethanol extract form at the same flavonoid inclusion level.

Serum protein expression in Equine Glandular Gastric Disease (EGGD) induced by phenylbutazone.

Equine Glandular Gastric Disease (EGGD) is a common disease in sport horses. This disease might be associated with usage of nonsteroidal anti-inflammatory drugs (NSAIDs) for treating inflammatory diseases. Although gastroscopy has been an effective method for diagnosis, but a less invasive, and inexpensive method is preferred. This study used proteomic technology to identify candidate serum proteins that might be used as markers of NSAIDs induced EGGD. Five Thoroughbred horses were given high doses of NSAID, phenylbutazone to treat lameness. The experiment was divided into three periods: (i) Pre-EGGD period, (ii) during EGGD period, and (iii) Post-EGGD period. Gastroscopy were used to diagnose EGGD, serum was collected to perform gel electrophoresis (1D SDS-PAGE) and mass spectrometry (LC-MS) in order to identify serum proteins in each group. The candidate serum proteins were computationally predicted for the interaction between phenylbutazone and proteins, tissue specific expression, and association to gastric ulceration. After EGGD induction, all horses showed clinical signs of colic with marked congestion and erosion appearing in the mucosa of the glandular stomach whereas no change was observed in the mucosa of non-glandular stomach. Our proteomic results identified 14 proteins that might be used as EGGD markers. These proteins were highly expressed in the glandular stomach and some proteins were associated with phenylbutazone or ulcer development. However, confirmation of these candidate marker proteins is required with specific antibodies in the larger horse population before they can be considered for application in the field.

Effect of Lactobacillus salivarius on growth performance, diarrhea incidence, fecal bacterial population and intestinal morphology of suckling pigs challenged with F4+ enterotoxigenic Escherichia coli.

OBJECTIVE: Gut health improvements were monitored with respect to growth performance, diarrhea incidence, fecal bacterial population and intestinal morphology of suckling pigs orally supplemented with live Lactobacillus salivarius (L. salivarius) oral suspensions and challenged with F4+ enterotoxigenic Escherichia coli (ETEC). METHODS: Two groups of newborn pigs from 18 multiparous sows were randomly designated as non-supplemented (control: n = 114 piglets) and L. salivarius supplemented groups (treatment: n = 87 piglets). Treatment pigs were orally administered with 2 mL of 109 colony-forming unit (CFU)/mL L. salivarius on days 1 to 3, then they were orally administered with 5 mL of 109 CFU/mL L. salivarius on days 4 to 10, while those in control group received an equal amount of phosphate buffered saline solution. On day 24 (2 weeks post supplementation), one pig per replicate of both groups was orally administered with 108 CFU/mL F4+ ETEC, then they were euthanized on day 29 of experiment. RESULTS: Results revealed that pigs in treatment group had a statistically significant increase in average daily gain, body weight and weight gain, and tended to lower diarrhea throughout the study. Numbers of Lactobacillus population in feces of treatment pigs were higher than control pigs, especially on day 10 of study. Numbers of total bacteria in intestinal contents of control pigs were also increased, but not Coliform and Lactobacillus populations. Histological examination revealed statistically significant improvements of villous height and villous/crypt ratio of duodenum, proximal jejunum and distal jejunum parts of treatment pigs compared with controls. Duodenal pH of treatment group was significantly decreased. CONCLUSION: Oral supplementation of live L. salivarius during the first 10 days of suckling pig promoted growth performance and gut health, reduced diarrhea incidence, increased fecal Lactobacillus populations and improved intestinal morphology.