Ixodid ticks from wild and domestic animals in East and Central Asian flyways.

Tick-borne diseases have a significant impact on human and animal populations, posing an increasing threat to public health, particularly in the context of climate change. Along with the various natural hosts of ticks, birds play a notable role in transmitting ticks and tick-borne pathogens, indicating the importance of monitoring flyways and establishing a cooperative network for comprehensive surveillance and to collect diverse tick samples across various regions. This study aimed to develop an international network for surveillance of disease, collection of sufficient tick samples, and overall identification of the geographical distribution of host and ticks in Asian regions, especially in 11 countries on East Asian and Central Asian flyways. Ticks were collected from wild animals, domestic animals, and vegetation to identify the differences between Ixodid ticks and understand tick distribution. We collected a total 6,624 of ticks from 11 collaborating Asian countries, the Republic of Korea (ROK), Japan, Thailand, Philippines, Indonesia, Cambodia, Vietnam, Taiwan, Hong Kong, Mongolia and Pakistan. We identified 17 host animals and 47 species of both residential and migratory birds. Ticks from birds collected from four countries (ROK, Japan, Hong Kong and Mongolia) belonged to two genera, Haemaphysalis and Ixodes, including Haemaphysalis (H.) longicornis, H. flava, H. concinna, H. hystricis, H. formosensis, Ixodes (I.) nipponensis and I. persulcatus. The potential of migratory birds to cross ecological barriers with ticks and tick-borne diseases indicated the need for further investigations to understand the migration of birds as potential vectors and the new influx of zoonotic diseases along migratory bird flyways. This study suggests the potential risk of spreading tick-borne diseases through birds, thus highlighting the importance of international cooperative networking.

The impact of urine storage methods on the results of quantitative bacterial culture in dog and cat.

Quantitative bacterial culture (QBC) is the gold standard for determining urinary tract infections. However, the majority of urine samples were sent to a lab for further bacterial culture. Therefore, the storage condition was vital to maintain the quality and accuracy of the urine samples. The main objective of our study was to examine the urine QBC outcomes of (1) immediate culture, (2) culture after urine sample storage at ambient temperature for 24 h, and (3) culture after urine sample storage at 4 °C for 24 hours. There were 49 feline samples and 30 canine samples included in this study. All QBC samples kept at ambient temperature and refrigerator were consistent with immediate QBC in cats. Eight positive results from immediate QBC were in accordance with refrigerated results in dogs. There were ten positive results in the room-temperature sample with two false-positive results. Our study showed that storing conditions at room temperature or refrigeration for 24 h does not impact the results of QBC in cat urine samples. For dog samples, chilled samples have a higher accuracy rate than room temperature samples, although the overall agreement was still satisfactory.

Paired Analysis of D-Dimer and Its Correlated Hemostatic Parameters in 30 Dogs with Neoplasms after Tumorectomy.

Previous studies have reported that dogs with neoplasms had elevated D-dimer levels. However, few studies have addressed whether D-dimer could be an indicator of tumor burden. The clinical significance of paired analysis of pre- and post-operation of D-dimer levels in dogs has rarely been described. The present study investigated the values of D-dimer levels and their correlated hemostatic alterations in dogs with surgically removable benign and malignant tumors. This study analyzed 30 clinically healthy and 30 tumor-bearing dogs and evaluated the hemostatic functions including D-dimer, thromboelastography G (TEG G), fibrinogen, activated partial thromboplastin time (aPTT), prothrombin time, and platelet count. The median level of pre-treatment D-dimer was 0.8 µg/mL (range: 0.1-6.3 µg/mL), whereas the control dogs exhibited a median value of 0.1 µg/mL (range: 0.1-0.1 µg/mL, p < 0.0001). After tumorectomy, the median levels of D-dimer (p < 0.0001), fibrinogen (p < 0.0001), TEG G value (p < 0.01), and aPTT (p < 0.05) were significantly lower than those of the pre-treatment samples. However, further studies are needed to clarify the values of other hemostatic evaluations. The study revealed the clinical significance of D-dimer and its correlated hemostatic parameters by paired analysis in dogs with tumors. Though more cases are needed for solid confirmation, these values could be potential tumor biomarkers for dogs.

Investigation of Toxoplasma infection in zoo animals using multispecies ELISA and GRA7 nested PCR.

BACKGROUND: Toxoplasma is an obligate intracellular protozoan that causes an important zoonotic disease with a worldwide distribution. Felids are the definitive hosts of this parasite, while virtually all warm-blooded animals, including birds, serve as intermediate hosts. Four ring-tailed lemurs (Lemur catta) in the Taipei Zoo died of acute Toxoplasma infection in June 2019. Since then, Toxoplasma has occasionally been identified in this Zoo during necropsy of dead animals and PCR of animal blood samples. Therefore, a general survey of Toxoplasma infection in animals in the Zoo seems to be needed. METHODS AND RESULTS: An indirect multispecies ELISA was used for the first time to screen for Toxoplasma infection in 326 serum samples collected from 75 species of animals. The infection rate of Toxoplasma was 27% (88/326). A commercial latex agglutination (LAT) assay was used to re-examine the samples with doubtful and uncertain ELISA results (151 samples from 42 species). The infection rate increased to 36.2% (118/326), and the indirect multispecies ELISA appeared to be applicable to 31 of 75 species animals included in this study. Nested PCR assays targeting the dense granule protein 7 (GRA7) gene and B1 gene were also used to detect Toxoplasma in DNA samples extracted from 10 liver or blood specimens from 8 animals. GRA7 gene fragments were amplified from 8 samples from 7 animals, while B1 gene fragments were amplified from only 4 samples from 4 animals. From the B1 nested PCR and the sequence data of GRA7 fragments amplified from infectious specimens, the animals in the Zoo were speculated to have been infected by at least three different Toxoplasma variants. CONCLUSIONS: According to the serological investigation, we speculated that over one-third (36.2%) of animals in Taipei Zoo presented the infection of Toxoplasma, and the indirect multispecies ELISA we used can be applied to detect Toxoplasma infection in 31 animal species included in this study. Sequence analysis revealed that at least three Toxoplasma variants were infecting the animals of Taipei Zoo.

The High Expression of Legumain in Canine Neoplasms: A Retrospective Analysis of 100 Cases.

Legumain, a novel asparaginyl endopeptidase, has been observed to be overexpressed in several types of human solid tumors. Elevated levels of legumain are found in human cancers, and this oncoprotein may facilitate tumor invasion and metastasis when overexpressed. These findings suggest that legumain plays a malignant role in cancer biology. However, currently, no publications have identified the role of legumain in the development of canine cancers. The present study first compared the expression patterns of legumain in paraffin-embedded canine tumor tissues, with those of normal tissues, by immunohistochemistry. A total of 100 canine tumor samples, including mast cell tumors, soft tissue sarcoma, hemangiosarcoma, lymphoma, mammary gland carcinoma, hepatoid gland tumor, squamous cell carcinoma, trichoblastoma, and melanoma were evaluated. Compared with the normal tissues, all tumor samples displayed high intensities of legumain expression. Mesenchymal-type tumors displayed immunoreactivity for legumain, with an average expression of 40.07% ± 1.70%, which was significantly lower than those of epithelial tumors and other types of tumors, which had median expressions of 49.12% ± 1.75% and 47.35% ± 2.71%, respectively (p < 0.05). These findings indicate that legumain has a high potential to be a candidate for distinguishing tumors from normal tissues. Although further studies on a larger number of cases are necessary to clarify the clinical application of legumain, the overexpression patterns of legumain in canine tumor tissues are reported, for the first time, in this study.

A molecular and serological survey in Taiwan to determine the true risk of babesiosis in dogs not receiving regular tick prevention.

Canine babesiosis is an important tick-borne disease worldwide. The prevalence varies between regions and countries; however, the incidence of tick infection is associated with the status of preventive tick control measures by the owner. To date, no studies have investigated the incidence of canine babesiosis and the condition of tick prevention in Taiwan. Therefore, the true risk of babesiosis could be underestimated in dogs that are not receiving tick prophylaxis. Samples were collected at 51 hospitals around Taiwan from 265 dogs not receiving regular tick prophylaxis. Diagnostic real-time PCR was performed, and 28 dogs (10.6%) were positive for Babesia spp., including B. gibsoni (26/28) and B. vogeli (2/28). Thirty-nine dogs (14.7%) were seropositive to B. gibsoni. Take the real-time PCR positive as the Babesia infected case, the positive and negative predictive value of serological assay were 64.1% and 98.7%, respectively. The seropositivity of B. gibsoni was significantly associated with real-time PCR positivity for Babesia spp. and vice versa (p < 0.001). The odds of seropositive representing real-time PCR positivity was 132.7 times greater than the seronegative (OR: 132.731, 95% CI 35.683-493.728). Risk factors in the population identified included: dogs with a short-haired coat; intact dogs; dogs from multi-dog households; dogs with more than 10 ticks and fleas on the skin; dogs that go outdoors more than 9 times per week; and dogs with an abnormal blood test result that included anemia, thrombocytopenia, and leukopenia. However, the dogs were not tested for other co-infections, therefore, these hematological risk factors should be carefully interpreted and confirmed by further diagnostic tests. In conclusion, when dogs present with abnormal blood test results and share the risk factors listed above, babesiosis should be seriously considered and followed up with molecular and serological testing. The serological assay used in this study can provide valuable information in diagnosing babesiosis in dogs in Taiwan.

L-Asparaginase, Doxorubicin, Vincristine, and Prednisolone (LHOP) Chemotherapy as a First-Line Treatment for Dogs with Multicentric Lymphoma.

Cyclophosphamide exhibits the weakest therapeutic effect compared with vincristine and doxorubicin in the CHOP (C, cyclophosphamide; H, doxorubicin; O, vincristine; and P, prednisolone) chemotherapeutic protocol for the treatment of canine lymphoma. Twenty dogs with multicentric lymphoma were treated using the LHOP protocol, which used l-asparaginase in place of cyclophosphamide, and the outcomes were historically compared with those of dogs that received CHOP chemotherapy in the same institution. No significant differences were found in age (p = 0.107), body weight (p = 0.051), sex (p = 0.453), clinical stage V (p = 1), substage b (p = 0.573), T-cell phenotype (p = 0.340), overall response (p = 1), and hypercalcaemia status (p = 1) between the LHOP and CHOP groups. The adverse effects of l-asparaginase were well tolerated and self-limiting. The median PFS (progression-free survival) and median ST (survival time) in the LHOP group were 344 days (range: 28-940 days) and 344 days (range: 70-940 days), respectively. The median PFS and median ST in the CHOP group were 234 days (range: 49-1822 days) and 314 days (range: 50-1822 days), respectively. The dogs that received LHOP chemotherapy had a significantly longer PFS than the dogs that received CHOP chemotherapy (p = 0.001). No significant difference was observed in ST between the LHOP and CHOP groups (p = 0.131). Our study findings thus indicate that the LHOP protocol can be used as a first-line chemotherapeutic protocol in canine multicentric lymphoma.

Association between weight change during initial chemotherapy and clinical outcome in dogs with multicentric lymphoma.

The majority of the known prognostic factors in dogs with lymphoma have been evaluated before treatment commences or at the time of diagnosis. Prognostic factors evaluated during the initial phase of treatment are less described but may provide important clinical information. In this retrospective study, 82 canine lymphoma patients were categorized according to the weight change between diagnosis and after 5 weeks of chemotherapy. Dogs that gained greater than 5% or lost greater than 5% of initial body weight were categorized as increased- or decreased-weight groups, respectively. Those in which weight changed less than 5% were categorized as the maintained-weight group. The median progression-free survival (PFS) in the increased-weight group, maintained-weight group and decreased-weight group was 226, 256 and 129 days, respectively. The decreased-weight group had significantly shorter PFS than the increased and maintained groups (P = .023, P = .003, respectively). The median survival time (ST) in the increased-weight group, maintained-weight group and decreased-weight group was 320, 339 and 222 days, respectively. There was no significant difference in ST among the three groups (P = .128). In Cox-regression results, weight change group and initial body weight were significant risk factors associated to PFS (P = .007, P = .001, respectively) while only patient's initial body weight was a significant risk factor to ST (P = .013). In conclusion, evaluation of initial body weight and weight changes over time can provide valuable information regarding PFS and ST in dogs with multicentric lymphoma.

Comparison of Simultaneous Measurements of Forelimb and Hindlimb Systolic Blood Pressure in Conscious Dogs.

Blood pressure is commonly measured in veterinary medicine. Previous studies have used indirect measurements to compare the forelimb and hindlimb blood pressure in dogs, and yielded contradictory results. Notably in these studies, the measurement of both limbs was not performed simultaneously. In this study, Doppler ultrasonography was used to measure systolic arterial pressure (SAP) and pulse rate (PR) simultaneously on the median palmar artery of forelimb and the medial plantar artery of hindlimb in 108 conscious dogs. The optimal cuff size was selected using 30-40% of the limb circumference. The SAP of forelimb and hindlimb were 149.5 ± 28.5 mm Hg and 152.3 ± 31.1 mm Hg. The PR of forelimb and hindlimb were 113.9 ± 31.7 beats per minute (bpm) and 113.1 ± 31.2 bpm, respectively. The SAP and PR of the forelimb and hindlimb correlated significantly (Pearson R2 = 0.696, P < .001; Pearson R2 = 0.966, P < .001). There was no significant difference in SAP and PR between the forelimb and hindlimb (P = .1; P = .184). There was no significant difference in the SAP and PR of the forelimb and hindlimb using Doppler device measurement in conscious dogs. Both sites of blood pressure measurement could be adequate for analyzing trends in blood pressure.

Outcome of Canine Multicentric Lymphoma after Single or Divided Treatment with Cyclophosphamide in Multidrug Chemotherapy.

Cyclophosphamide is commonly used in combination chemotherapy to treat dogs with lymphoma. The metabolite of cyclophosphamide, acrolein, can irritate urinary bladder and cause sterile hemorrhagic cystitis. Dividing the administration of cyclophosphamide across multiple days may reduce the concentration of this metabolite in urinary bladder and reduce the possibility of cystitis. However, the impact of the therapeutic effect of this modification is not evaluated and compared to traditional single maximum-tolerated dose regimen. Seventy-two dogs with multicentric lymphoma received either bolus doses or divided doses of cyclophosphamide were included in this study. The incidence of hemorrhagic cystitis between 2 cyclophosphamide treatment groups was not significantly different (P = .357). There was no statistical difference in progression-free survival and survival time between 2 groups (P = .267 and P = .346). This modification of cyclophosphamide administration did not reduce the side effect of cystitis or affect remission and survival times in lymphoma dogs.

Elevated plasma YKL-40 level is found in the dogs with cancer and is related to poor prognosis.

YKL-40, a secreted glycoprotein, may serve as an autoantigen, which mediates multiple inflammatory diseases and cancers. A high YKL-40 serum level is correlated with metastasis and poor survival in a variety of human cancers. However, the role of YKL-40 in dogs is still under evaluation. Herein, we examined the associations between plasma YKL-40 level and YKL-40 autoantibody (YAA) titers with malignancy and prognosis in canine cancer. Plasma levels of YKL-40 in healthy dogs (n = 20) and in dogs (n = 82) with cancer were evaluated using enzyme-linked immunosorbent assay. Our results indicated that plasma YKL-40 levels were significantly higher (p < 0.01) in dogs with cancer than in healthy dogs. A significant decrease in the YAA titers was detected in the dogs with cancer when compared with those of the healthy dogs (p < 0.05), although the change was not correlated with the YKL-40 levels. Among the dogs with cancer, plasma YKL-40 levels in the dogs that later relapsed or had metastasis were significantly higher than in the dogs with no signs of relapse (p < 0.01) or metastasis (p < 0.05). The relapse and metastasis rates were significantly higher in the high YKL-40 group (> 180 pg/mL) than in the low YKL-40 group (< 180 pg/mL). The results imply that plasma YKL-40 levels might have the potential to be developed as a marker of malignancy progression and prognosis in canine cancers.

The investigation of histopathology and locations of excised eyelid masses in dogs.

BACKGROUND: Eyelid masses are the most common ocular diseases in dogs. However, there are no studies specifically investigating the location of eyelid masses. METHODS: 118 dogs with 119 eyelid masses were included. Medical records and pathology reports were retrospectively reviewed at National Taiwan University veterinary hospital and Vision Eyecare Centre for Animals between 2012 and 2017. RESULTS: Mean age of dogs was 9.4±2.5 years. Female spayed dogs had significantly higher prevalence of eyelid mass than female intact dogs (p<0.01). Prevalence of upper eyelid mass was significantly higher than lower eyelid mass (p<0.01). The upper lateral eyelid was the significantly more common location compared with the upper medial eyelid. Mean mass volume was 258.2±661.0 mm3. The most common eyelid mass type was meibomian epithelioma (34.5%), followed by meibomian adenoma (29.4%) and meibomian hyperplasia (18.5%). Non-tumours comprised 25.2%, benign tumours comprised 67.2% and malignant tumours comprised 7.6% of all eyelid masses. Malignant tumours were significantly larger than benign lesions (p<0.01). All dogs underwent surgical excision without any complications. CONCLUSION: The majority of eyelid masses were benign. Surgical intervention can prevent further ocular irritation and provide good prognosis.

Assessment of temporal association of relapse of canine multicentric lymphoma with components of the CHOP protocol: Is cyclophosphamide the weakest link?

Combination chemotherapy, using cyclophosphamide, hydroxydaunorubicin, vincristine and prednisolone (CHOP), is the most commonly used treatment for canine lymphoma. Most affected dogs respond during the initial stages of chemotherapy, but many relapse. The aim of this study was to evaluate the relationship between the use of specific chemotherapy drugs and clinical relapse, using the modified Madison-Wisconsin, 25 week chemotherapy protocol. Forty-one of 68 dogs affected with multicentric lymphoma relapsed during the treatment period. Relapse occurred more frequently after the administration of cyclophosphamide (n = 24; P < 0.01), compared with vincristine (n = 9) or doxorubicin (n = 5). Therefore, the therapeutic outcome of traditional CHOP-based chemotherapy might be improved by replacing cyclophosphamide with other cytotoxic drugs.

Comparison of efficacy and toxicity of doxorubicin and mitoxantrone in combination chemotherapy for canine lymphoma.

Forty-four dogs with multicentric lymphoma were treated using a cyclophosphamide, doxorubicin, vincristine, and prednisolone (CHOP) induction protocol or treated using a cyclophosphamide, mitoxantrone, vincristine, and prednisolone (CMOP) induction protocol. There was no statistical difference in signalment and the presence of historical negative prognostic factors between the groups. The median progression-free survival (PFS) in the CHOP and CMOP groups were 222 d and 162 d, respectively (P = 0.75). The median survival time (MST) of dogs in CHOP and CMOP groups were 318 d and 242 d, respectively (P = 0.63). Anorexia and diarrhea episodes were significantly higher in the CHOP group than in the CMOP group (P = 0.02 and P = 0.01, respectively). These results suggest that the CMOP protocol provides similar PFS, MST and causes fewer side effects compared to the CHOP protocol. Therefore, the CMOP protocol may be another treatment choice for canine multicentric lymphoma.

Comparaison de l'efficacité et de la toxicité de la doxorubicine et du mitoxantrone dans la chimiothérapie combinée pour le lymphome canin. Quarante-quatre chiens atteints d'un lymphome multicentrique ont été traités à l'aide d'un protocole d'induction au cyclophosphamide, à la doxorubicine, à la vincristine et à la prednisolone (CHOP) ou traités en utilisant un protocole d'induction à la cyclophosphamide, au mitoxantrone, à la vincristine et à la prednisolone (CMOP). Il n'y a eu aucune différence statistique dans le signalement et la présence des facteurs de pronostic négatifs historiques entre les groupes. La survie sans progression (SSP) médiane des groupes CHOP et CMOP était de 222 jours et de 162 jours, respectivement (P = 0,75). La durée de survie moyenne (DSM) des chiens des groupes CHOP et CMOP a été de 318 jours et de 242 jours, respectivement (P = 0,63). Les épisodes d'anorexie et de diarrhée ont été significativement plus élevés dans le groupe CHOP comparativement au groupe CMOP (P = 0,02 et P = 0,01, respectivement). Ces résultats suggèrent que le protocole CMOP offre une SSP et une DSM semblables tout en causant moins d'effets secondaires comparativement au protocole CHOP. Par conséquent, le protocole CMOP peut représenter un autre choix de traitement pour les lymphomes multicentriques canins.(Traduit par Isabelle Vallières).