External Hoof Measurements of Untrimmed and Unshod Mules in Northern Thailand.

External hoof characteristics, balance, and conformation have been extensively studied in horses; however, mules remain understudied in these aspects. This study evaluated the size, shape, and symmetry of untrimmed and unshod forelimb hooves, compared the symmetry between forelimb hooves and stratified external forelimb hoof measurements based on the body condition score of mules raised in the foothill plains of northern Thailand. The forelimb hooves of 38 mules were photographed and 33 parameters, including angular and linear measurements, were analyzed. A multivariate analysis was used to explore the influence of sex, age, and body condition scores (BCS) on angular, linear, and area parameters. Additionally, one-way ANOVA followed by Tukey's test was used to compare these parameters across different BCS groups. Despite the absence of shoeing and trimming, these mules exhibited optimal left-right forelimb hoof symmetry, with no significant (p < 0.05) differences in: outer wall length and inner wall length (OWL-IWL: Left 0.11 ± 0.66 cm; Right -0.12 ± 0.43 cm); sole length and sole width (SLS-SW: Left 1.65 ± 0.76 cm; Right 1.46 ± 0.89 cm); dorsal hoof wall length and heel length (DHWL-HL: Left 4.00 ± 0.80 cm; Right 3.81 ± 0.72 cm); and frog length and frog width (FL-FW: Left 3.88 ± 1.13 cm; Right 3.82 ± 0.18 cm). However, significant (p < 0.05) differences were observed within each body condition score group for forelimb hoof measurements for DHWL, IWL, heel separation (HS), heel bulb distance (HBD), SW, FW, and FL, while sex and age had no significant differences across the study variables. These findings provide valuable insights into mule welfare and management, contributing to understanding of the interplay between overall health and hoof conformation in the study area.

Immunological Cross-Reactivity of Proteins Extracted from the Oomycete Pythium insidiosum and the Fungus Basidiobolus ranarum Compromises the Detection Specificity of Immunodiagnostic Assays for Pythiosis.

Pythiosis, a life-threatening disease caused by Pythium insidiosum, has been increasingly diagnosed worldwide. A recently developed immunochromatographic test (ICT) enables the rapid diagnosis of pythiosis. During the 3-year clinical implementation of ICT in Thailand, we collected the laboratory reports of 38 animals with suspected pythiosis and detected ICT false-positive results in three horses and a dog with basidiobolomycosis. P. insidiosum and Basidiobolus ranarum cause infections with indistinguishable clinical and microscopic features. This study investigated cross-reactive antibodies by probing P. insidiosum and B. ranarum crude extracts and cell-free synthesized I06 protein (encoded in P. insidiosum genome, not other fungi) against a panel of pythiosis, basidiobolomycosis, rabbit anti-I06 peptide, and control sera by Western blot analyses. ICT false-positive results occurred from the cross-reactivity of anti-B. ranarum antibodies to the 15, 50, 60, and 120 kDa proteins of P. insidiosum, not double infections caused by both pathogens. Notably, ICT could help to screen pythiosis, and the positive test requires confirmation by culture or molecular method. The detection specificity of ICT requires improvement. The crude extract containing multispecies antigens needs replacement with a refined P. insidiosum-specific protein. We proposed that the 55 kDa I06 protein is an excellent candidate for developing a more specific serodiagnostic test for pythiosis.

Modified Z-bar shoe eliminates occasional frog bruising accompanying Z-bar shoeing for navicular syndrome management in underrun-heeled horses.

Two horses diagnosed with underrun heels leading to navicular syndrome were fitted with Z-bar shoes. They occasionally showed moderate lameness on the affected legs after the sixth and tenth consecutive farrier adjustments. The affected hooves were sensitive to compression of the frog and sole in their palmar regions. Modified Z-bar shoes were created and shod on the lame legs based on previous with regional anaesthesia and recent clinical examination. The lameness scores were slightly improved at the first shoeing with the modified Z-bar shoes but were markedly improved at four and eight weeks aftershoeing, without medication. The modified Z-bar shoes in this report could potentially protect the palmar structure damaged by usual Z-bar shoes and contribute to reducing pathogenic impacts deriving from underrun heels and navicular syndrome.

Identification and Biotyping of Pythium insidiosum Isolated from Urban and Rural Areas of Thailand by Multiplex PCR, DNA Barcode, and Proteomic Analyses.

Pythium insidiosum causes pythiosis, a fatal infectious disease of humans and animals worldwide. Prompt diagnosis and treatment are essential to improve the clinical outcome of pythiosis. Diagnosis of P. insidiosum relies on immunological, molecular, and proteomic assays. The main treatment of pythiosis aims to surgically remove all affected tissue to prevent recurrent infection. Due to the marked increase in case reports, pythiosis has become a public health concern. Thailand is an endemic area of human pythiosis. To obtain a complete picture of how the pathogen circulates in the environment, we surveyed the presence of P. insidiosum in urban (Bangkok) and rural areas of Thailand. We employed the hair-baiting technique to screen for P. insidiosum in 500 water samples. Twenty-seven culture-positive samples were identified as P. insidiosum by multiplex PCR, multi-DNA barcode (rDNA, cox1, cox2), and mass spectrometric analyses. These environmental strains of P. insidiosum fell into Clade-II and -III genotypes and exhibited a close phylogenetic/proteomic relationship with Thai clinical strains. Biodiversity of the environmental strains also existed in a local habitat. In conclusion, P. insidiosum is widespread in Thailand. A better understanding of the ecological niche of P. insidiosum could lead to the effective prevention and control of this pathogen.

Protein A/G-based enzyme-linked immunosorbent assay for detection of anti-Pythium insidiosum antibodies in human and animal subjects.

OBJECTIVES: Pythiosis is a deadly infectious disease caused by Pythium insidiosum. Reports of both human and animal pythiosis are on the rise worldwide. Prognosis of the pythiosis patients relies on early diagnosis and prompt treatment. There are needs for an immunodiagnostic test that can detect the disease in both humans and animals. This study aims at reporting an optimized protocol for the development of a protein A/G-based enzyme-linked immunosorbent assay (ELISA) for the detection of anti-P. insidiosum antibody in multiple host species. RESULTS: A total of 25 pythiosis and 50 control sera, obtained from humans, horses, dogs, cats, and cows, were recruited for the assay development. With a proper ELISA cutoff point, all pythiosis sera can ultimately be distinguished from the control sera. The successfully-developed protein A/G-based ELISA can detect the anti-P. insidiosum antibodies in serum samples of both humans and animals. It is a versatile, feasible-to-develop, and functional immunodiagnostic assay for pythiosis.

First confirmed case of nasal pythiosis in a horse in Thailand.

INTRODUCTION: Pythiosis is caused by Pythium insidiosum, a fungus-like organism in the class Oomycetes. It can infect humans and a variety of animal species in tropical, subtropical and some temperate regions. Cases of animal pythiosis have occurred predominantly in horses in the skin and subcutaneous tissue at the limbs and in the ventral portion of thoracoabdominal wall - lesions in the nasal region are rarely reported. Moreover, although many human pythiosis cases have been reported in Thailand, no cases of animal pythiosis in Thailand have been reported. CASE PRESENTATION: We report a case of pythiosis in a horse infected at the nasal cavity. Diagnosis was performed by zoospore formation by bait technique, immunohistochemical stain, immunochromatography and sequence analysis. CONCLUSION: The sequences of rDNA were 99 % and 96 to 99 % identical to GenBank isolates of Pythium insidiosum from two Thai human patients and horses from various countries, respectively. This represents the first confirmed report of nasal equine pythiosis in Thailand.