Effect of feeding toy and the presence of a dog owner during the feeding time on dog welfare.

Background and Aim: A conventional feeding bowl is the primary method that dog owners use to feed their dogs, but this may not encourage natural behaviors and may even exacerbate unwanted behaviors. This study aimed to compare a conventional feeding bowl to a feeding toy in relation to behavior, cortisol levels, and heart rate variability (HRV). Materials and Methods: The behaviors of four dogs were recorded and analyzed while being fed using either a stainless bowl (B) or a feeding toy (T) and either alone (A) or accompanied by a dog owner (O) for 30 min with each treatment (BA, BO, TA, and TO treatments). The dogs that were fed alone with the stainless bowl (BC) or the feeding toy (TC) were fed for 15 min/day for 7 days with their treatment, and serum cortisol levels measured on the first and last days of treatment. The dogs fed by the stainless bowl (BH) or the feeding toy (TH) with the owner present for 15 min for each treatment had their heart rate (HR) and HRV recorded by Polar® H10 during feedings The results were compared using a one-way analysis of variance (ANOVA), repeated measure ANOVA, and Student's t-test. Results: The dogs spent more time eating and interacting with the feeding toys than stainless bowls. The activity of the dogs was higher when using feeding toys, particularly with the TO treatment. Cortisol levels were significantly lower on day 7 than on day 1 of the TC treatment. The dogs' HR was higher during TH treatment than during BH treatment. All HRV parameters were decreased significantly when feeding the dog with the toys. Conclusion: The results of this study support the idea that feeding enrichment supports the natural feeding behaviors of dogs as they mimic hunting and playing behaviors. This reduced unwanted behavior, cortisol levels, and HRV, and increased food consumption, eating duration, and active behaviors. The presence of the dog's owner is important because it can enhance feeding and active behaviors, and feeding enrichment can improve the dog's welfare and the dog-human relationship.

Factors associated with pet dog behavior in Thailand.

Background and Aim: Dog behavior problems pose serious public health and economic and animal welfare concerns. There are many factors influencing dog behavior. This study aims to explore factors associated with pet dog behavior in Thailand using the Canine Behavioral Assessment and Research Questionnaire (C-BARQ). Materials and Methods: The Thai version of the C-BARQ was validated in 2022. The online C-BARQ survey (Google Forms) was advertised and distributed on social media for 3 months. There were a total of 1827 respondents to the survey. The relationship among 12 subscales and variables was analyzed using Spearman's correlation. Results: "Excitability," "attachment- and attention-seeking," and "chasing" were the three most reported behavior problems. "Trainability," a favored dog behavior, also had a high score in the study. These behaviors were associated with the owner's age, house type, the dog's historical background, the number of dogs and the presence of other species in the household, the dog's training, diet types, and the daily number of meals. The lowest mean score was for "owner-directed aggression," and it was associated with the dog's gender and size, the number of dogs and the presence of other species in the household, the dog's training, and the number of daily meals. Conclusion: This is the first empirical study demonstrating factors related to dog behavior in Thailand. It provides an in-depth understanding of the prevalence and factors associated with Thai pet dog behavior and important knowledge for further studies to advocate for dog-human relationships and contribute to a reduction in dog abandonment in Thailand.

Hematological analysis of naturally infecting blood parasites in dogs.

Background and Aim: Blood parasite infections such as anaplasmosis, babesiosis, and ehrlichiosis are commonly found in domestic dogs, which adversely influence their health. Many dogs are infected with multiple blood parasites that cause more severe diseases than a single infection. This study aimed to investigate the effect of multiple blood parasite infections on the hematological profiles of dogs at a shelter in Southern Thailand. Materials and Methods: The blood samples from 122 dogs were collected to assess the hematology profiles of uninfected, single-infected, and multiple blood parasite-infected dogs. The results were compared using Kruskal-Wallis test and Dwass-Steel-Critchlow-Fligner pairwise comparisons. The infections were confirmed by polymerase chain reaction. Results: The results showed that all the infected dogs had significantly lower red blood cell (RBC) count, hemoglobin (HB), hematocrit (HCT), and platelet count (PLT) than the uninfected dogs. Although the dogs with triple infection had lower RBC, HB, HCT, and PLT than those with double and single infections, the difference was not statistically significant. Conclusion: We proposed that triple blood parasite infection with Anaplasma platys, Babesia vogeli, and Ehrlichia canis caused more severe disease than double and single infections. Evaluating the hematological profiles of dogs naturally infected with single, double, and multiple blood parasite infections without clinical signs can enhance their health and welfare.

Gymnema inodorum Leaf Extract Improves Cardiac Function in Experimental Mice Infected with Plasmodium Berghei.

Malaria-associated cardiac injury has been reported to be the primary cause of death due to severe malaria. The discovery of substances showing a protective effect on cardiac injury during malaria infection is urgently needed. Hence, the purpose of this study was to evaluate the efficacy of Gymnema inodorum leaf extract (GIE) on cardiac function in mice infected with Plasmodium berghei. ICR mice were treated with 1 × 107 infected red blood cells of P. berghei ANKA (PbANKA), administered orally with GIE in 100, 250 and 500 mg/kg body weight of mice. Creatine phosphokinase (CPK) and echocardiography were carried out. It was found that CPK and heart-weight to body-weight (HW/BW) ratios were significantly higher in untreated mice than the healthy control. Moreover, impaired cardiac function in the untreated group was observed as indicated by changes in echocardiography. Interestingly, GIE exerted a protective effect on cardiac injury induced by PbANKA infection. Our results demonstrated that the parasitemia percentage, CPK, HW/BW ratio, and echocardiography in GIE treated mice were improved. However, there was no significant difference between GIE dosages. Therefore, GIE possessed a cardio-protective effect during malaria infection in mice.

First study on molecular detection of hemopathogens in tabanid flies (Diptera: Tabanidae) and cattle in Southern Thailand.

Background and Aim: Female tabanids play a key role in disease transmission as mechanical vectors for various hemopathogens, but only a limited number of studies have been conducted on them. This study aimed to investigate the occurrence of hemopathogens in tabanid flies compared to those found in nearby cattle hosts. Materials and Methods: Tabanids were collected using a Nzi trap for three consecutive days per month during the dry season (February-May 2021). Furthermore, blood samples were collected from 20 beef cattle (Bos taurus) raised in the same area where the flies were captured. Conventional polymerase chain reaction (PCR) was used to detect hemopathogenic DNA in flies and beef cattle. Results: In total, 279 female tabanids belonging to five species were collected: Tabanus megalops, Tabanus rubidus, Tabanus mesogaeus, Chrysops dispar, and Chrysops fuscomarginalis. Notably, T. megalops was the most abundant species, accounting for 89.2% of the flies collected (n = 249). PCR technique revealed that 76.6% of T. megalops carried at least one pathogen (Anaplasma, Ehrlichia, Babesia, or Theileria). In addition, all beef cattle had multiple hemopathogenic infections (Anaplasma marginale, Ehrlichia spp., Babesia bigemina, Babesia bovis, and Theileria spp.). Conclusion: Although T. megalops could carry many hemopathogens, it might not be an important vector due to the limited number of flies and parasitic load. Furthermore, T. megalops could be utilized to monitor the presence of hemopathogens in the study area, but not the disease occurrence in the individual host species. Knowing the presence of hemopathogens in flies could help manage the disease in this area.

Multiple blood pathogen infections in apparently healthy sheltered dogs in southern Thailand.

In developing countries such as Thailand, free-ranging dogs are frequently involved in road accidents and contribute to the cost of public healthcare. Shelters play a vital role in communities because they help to control the population of unwanted and free-ranging dogs. This study aimed to investigate blood pathogen infection in sheltered dogs, as it is one of the factors contributing to animal welfare. Blood samples were randomly collected from 141 dogs from the largest shelter (approximately 400-500 dogs in total) in southern Thailand. Blood pathogens were detected using both PCR and light microscopy. Four blood pathogens were identified: Anaplasma platys, Ehrlichia canis, Babesia canis vogeli, and Hepatozoon canis. No trypanosomes were detected. The incidence of blood parasite infection was 56.7% (80/141) by PCR, and 28.4% (40/141) by microscopy. E. canis was the most prevalent pathogen, accounting for 46.1% (65/141) of the cases, while multiple infections accounted for 22% (31/141) of the cases. A triple infection with E. canis, A. platys, and B. canis vogeli was observed in 5.7% (8/141) of the cases. Although PCR is far more sensitive than microscopy, it appears to have equivalent specificity. In conclusion, this study reported a high occurrence of blood pathogen infections in clinically healthy sheltered dogs. Many of them were infected with multiple pathogens and may have been infected before entering the shelter. These findings suggest that a blood test is necessary to screen dogs prior to their admission to the shelter to prevent disease transmission and enhance animal welfare.

Antimicrobial resistance in Staphylococcus pseudintermedius on the environmental surfaces of a recently constructed veterinary hospital in Southern Thailand.

Background and Aim: Staphylococcus pseudintermedius is a zoonotic bacterium commonly found in animals, especially dogs. These bacteria can survive on environmental surfaces for several months. The infection of S. pseudintermedius from the environment is possible, but properly cleaning surface objects can prevent it. This study aimed to investigate the prevalence of methicillin-resistant S. pseudintermedius (MRSP) in the environment of a recently constructed veterinary hospital in Southern Thailand, where we hypothesized that the prevalence of MRSP might be very low. Materials and Methods: At three different time points, 150 samples were collected from different environmental surfaces and wastewater across the veterinary hospital. The collection was done after the hospital's cleaning. Bacteria were purified in the culture before being identified as species by biochemical tests and polymerase chain reaction (PCR). Next, the antimicrobial-resistant profile was performed using an automated system (Vitek 2). Finally, the antimicrobial resistance genes were identified using PCR. Results: Fifteen colonies of S. pseudintermedius were isolated from the surfaces of eight floors, four tables, two chairs, and one rebreathing tube. Fourteen colonies (93.3%) were multidrug-resistant (MDR) and carried the blaZ gene (93.3%). The majority of colonies were resistant to benzylpenicillin (93.3%), cefovecin (93.3%), ceftiofur (93.3%), kanamycin (93.3%), and neomycin (93.3%). Notably, only four colonies (26.7%) were methicillin-susceptible S. pseudintermedius, whereas 11 colonies (73.3%) were MRSP and carried both the mecA and blaZ genes. Five MRSP (45.5%) were resistant to at least 14 antimicrobial drugs, represented as extensively drug-resistant (XDR) bacteria. Ten of eleven MRSP (90.9%) were Staphylococcal chromosomal mec type V, while another displayed untypeable. Despite the routine and extensive cleaning with detergent and disinfectant, MRSP isolates were still detectable. Conclusion: Many isolates of MRSP were found in this veterinary hospital. Almost all of them were MDR, and nearly half were XDR, posing a threat to animals and humans. In addition, the current hospital cleaning procedure proved ineffective. Future research should be conducted to determine the bacterial biofilm properties and bacterial sensitivity to certain detergents and disinfectants.

Suppression of cardiomyocyte functions by ß-CTX isolated from the Thai king cobra (Ophiophagus hannah) venom via an alternative method.

BACKGROUND: Beta-cardiotoxin (ß-CTX), the three-finger toxin isolated from king cobra (Ophiophagus hannah) venom, possesses ß-blocker activity as indicated by its negative chronotropy and its binding property to both ß-1 and ß-2 adrenergic receptors and has been proposed as a novel ß-blocker candidate. Previously, ß-CTX was isolated and purified by FPLC. Here, we present an alternative method to purify this toxin. In addition, we tested its cytotoxicity against different mammalian muscle cell types and determined the impact on cardiac function in isolated cardiac myocyte so as to provide insights into the pharmacological action of this protein. METHODS: ß-CTX was isolated from the crude venom of the Thai king cobra using reverse-phased and cation exchange HPLC. In vitro cellular viability MTT assays were performed on mouse myoblast (C2C12), rat smooth muscle (A7r5), and rat cardiac myoblast (H9c2) cells. Cell shortening and calcium transient dynamics were recorded on isolated rat cardiac myocytes over a range of ß-CTX concentration. RESULTS: Purified ß-CTX was recovered from crude venom (0.53% w/w). MTT assays revealed 50% cytotoxicity on A7r5 cells at 9.41 ± 1.14 µM (n = 3), but no cytotoxicity on C2C12 and H9c2 cells up to 114.09 µM. ß-CTX suppressed the extend of rat cardiac cell shortening in a dose-dependent manner; the half-maximal inhibition concentration was 95.97 ± 50.10 nM (n = 3). In addition, the rates of cell shortening and re-lengthening were decreased in ß-CTX treated myocytes concomitant with a prolongation of the intracellular calcium transient decay, indicating depression of cardiac contractility secondary to altered cardiac calcium homeostasis. CONCLUSION: We present an alternative purification method for ß-CTX from king cobra venom. We reveal cytotoxicity towards smooth muscle and depression of cardiac contractility by this protein. These data are useful to aid future development of pharmacological agents derived from ß-CTX.

Suppression of cardiomyocyte functions by ß-CTX isolated from the Thai king cobra (Ophiophagus hannah) venom via an alternative method

Beta-cardiotoxin (ß-CTX), the three-finger toxin isolated from king cobra (Ophiophagus hannah) venom, possesses ß-blocker activity as indicated by its negative chronotropy and its binding property to both ß-1 and ß-2 adrenergic receptors and has been proposed as a novel ß-blocker candidate. Previously, ß-CTX was isolated and purified by FPLC. Here, we present an alternative method to purify this toxin. In addition, we tested its cytotoxicity against different mammalian muscle cell types and determined the impact on cardiac function in isolated cardiac myocyte so as to provide insights into the pharmacological action of this protein. Methods: ß-CTX was isolated from the crude venom of the Thai king cobra using reverse-phased and cation exchange HPLC. In vitro cellular viability MTT assays were performed on mouse myoblast (C2C12), rat smooth muscle (A7r5), and rat cardiac myoblast (H9c2) cells. Cell shortening and calcium transient dynamics were recorded on isolated rat cardiac myocytes over a range of ß-CTX concentration. Results: Purified ß-CTX was recovered from crude venom (0.53% w/w). MTT assays revealed 50% cytotoxicity on A7r5 cells at 9.41 ± 1.14 µM (n = 3), but no cytotoxicity on C2C12 and H9c2 cells up to 114.09 µM. ß-CTX suppressed the extend of rat cardiac cell shortening in a dose-dependent manner; the half-maximal inhibition concentration was 95.97 ± 50.10 nM (n = 3). In addition, the rates of cell shortening and re-lengthening were decreased in ß-CTX treated myocytes concomitant with a prolongation of the intracellular calcium transient decay, indicating depression of cardiac contractility secondary to altered cardiac calcium homeostasis. Conclusion: We present an alternative purification method for ß-CTX from king cobra venom. We reveal cytotoxicity towards smooth muscle and depression of cardiac contractility by this protein. These data are useful to aid future development of pharmacological agents derived from ß-CTX.(AU)

Comparative effects of amiodarone and dronedarone treatments on cardiac function in a rabbit model.

AIM: The objective of the study was to compare the effects of amiodarone (AM) and dronedarone (DR) on heart rate variability (HRV) and cardiac contractility in a rabbit model. MATERIALS AND METHODS: A total of 16 male New Zealand white rabbits were divided into two groups, treated either with AM or DR at incremental dosages of 50 mg/kg/day (AM50 and DR50) and 100 mg/kg/day (AM100 and DR100), orally administrated for 7 days. At the end of each period, electrocardiograms were recorded during consciousness and analyzed using the short-term time and frequency domains of HRV. Standard echocardiography and speckle-tracking echocardiography were studied during immobilization with xylazine and ketamine. RESULTS: The results showed that AM100 and DR100 significantly decreased heart rate, total power, low-frequency component, and low-to-high frequency ratio compared with baselines. Most echocardiogram parameters revealed no significant difference from baselines, except for the global circumferential plane strain rate and time to peak standard deviation of strain, which had statistical significances after treating with AM. CONCLUSION: Both AM and DR possess negative chronotropy and reduce HRV, which may be explained by their sympathetic suppression and calcium channel blocking activities. Theoretically, both antiarrhythmic drugs may also possess negative inotropy, but only AM is shown to have a negative inotropic effect and reduces cardiac dyssynchrony in this model.