Erythrocyte sedimentation rate in heartworm naturally infected dogs "with or without" Leishmania infantum seropositivity: an observational prospective study.

Canine heartworm disease by Dirofilaria immitis and canine leishmaniosis by Leishmania infantum (CanL) are both vector-borne diseases with frequently overlapping endemicity and able to trigger the acute phase response, being characterized by variations in acute phase proteins (APP). Recently, erythrocyte sedimentation rate (ESR), an indicator of inflammation, has gained attention in veterinary medicine, proving useful in several conditions that include CanL active forms in dogs. This study aims to evaluate ESR in heartworm-infected dogs, compare levels with heartworm-infected and L. infantum seropositive dogs as well as clinically healthy dogs, and assess correlations with other laboratory parameters. From October 2022 to January 2023, a prospective observational study was conducted enrolling heartworm-infected (Dirofilaria group) and heartworm-infected L. infantum seropositive (Dirofilaria/Leishmania group) animals subgrouped according to the CanL clinical form (Dirofilaria/Leishmania active and non-active groups). A group of clinically healthy dogs (control group) was also included. For each dog enrolled physical examination and laboratory tests (complete blood count, biochemical panel including APP, serum protein electrophoresis) were performed. Dirofilaria and Dirofilaria/Leishmania groups presented a significantly higher ESR level compared to healthy dogs. Dirofilaria/Leishmania active group had the highest ESR level among the groups considered. Dirofilaria/Leishmania non-active group had an ESR similar to the Dirofilaria group, but significantly higher and lower compared to the control and the Dirofilaria/Leishmania active group, respectively. A significant positive correlation between ESR and C-Reactive Protein has been found in all groups except for the Dirofilaria/Leishmania non-active group. In Dirofilaria/Leishmania active group a strong positive correlation between ESR and gamma globulins percentage as well as a strong negative correlation between ESR and albumin, albumin/globulins ratio were found. Overall, the ESR was confirmed to be an inflammation marker as well as a helpful disease index, being notably increased in heartworm-infected dogs affected by an active form of CanL.

Inflammatory Status and Chronic Kidney Disease in Cats: Old and New Inflammatory Markers-A Pilot Prospective Study.

This prospective study aimed to evaluate inflammatory status in cats affected by chronic kidney disease (CKD) at IRIS stages 2-4, using serum amyloid A (SAA) and the erythrocyte sedimentation rate (ESR) as inflammatory markers. Thirty-two cats with CKD and ten clinically healthy cats (i.e., control group) were enrolled. The recording of signalment data, complete physical examinations, and abdominal ultrasonography were performed for each animal. Additionally, ESR levels, complete blood count, clinical chemistry (including SAA determination), serum protein electrophoresis, and complete urinalysis were executed. This study's results showed that mean ESR and SAA concentrations in cats with CKD were statistically higher compared to those of the control group (p = 0.0005 and p = 0.007, respectively). The SAA concentration was significantly increased at IRIS stages 2, 3, and 4 compared to the control group. Meanwhile, the ESR was significantly higher in cats at IRIS stages 3 and 4 (p = 0.0003 and p = 0.0007, respectively), but not at IRIS stage 2, compared to the control group. These results provide evidence that feline CKD is associated with a systemic inflammatory status. Moreover, the rise in ESR appears to be more linked to advanced stages of the disease and could, therefore, correlate with the uremic condition.

Assessment of the diagnostic performance of serological tests in areas where Leishmania infantum and Leishmania tarentolae occur in sympatry.

BACKGROUND: Visceral leishmaniosis caused by infection with the zoonotic protozoan Leishmania infantum is a life-threatening disease affecting dogs and humans. The sympatric occurrence of L. infantum and Leishmania tarentolae in an area of southern Italy endemic for canine leishmaniosis, where dogs are also exposed to the latter species, suggests the persistence of herpetophilic L. tarentolae in a non-permissive host, therefore raising questions about the performance of serological diagnostic tests routinely employed. METHODS: The diagnostic performance of serological tests such as the immunofluorescence antibody test (IFAT), two commercial immunoenzymatic assays (i.e. NovaTec VetLine Leishmania ELISA® and rK39 ICT®) and an in-house enzyme-linked immunosorbent assay (ELISA) was evaluated in healthy dogs seropositive to L. infantum, whereas the only IFAT available was used to detect antibodies to L. tarentolae. RESULTS: With the IFAT, out of a total of 104 dogs tested, 15 were seronegative for L. infantum of which three were L. tarentolae seropositive' and 89 were L. infantum seropositive. Of the latter 89 dogs, representing the highest proportion of seropositive animals (85.6%) detected by IFAT' 66 were also seropositive for L. tarentolae. Cohen's kappa (κ) agreement coefficient between the IFAT results and those of all the other tests was very low, and the IFAT results were significantly different from those of all the other serological tests as calculated by Cochran's Q-test. Analysis using the Bayesian latent class (Bayes-LCA) showed that the in-house ELISA and IFAT contributed the most towards identifying infected and non-infected dogs, respectively. The IFAT test showed low positive predictive value (59.5%), but high negative predictive value (100%). CONCLUSIONS: These results demonstrate that the IFAT for L. infantum, although highly sensitive, may not be considered a useful diagnostic test due to its low specificity. Therefore, an accurate serological tool with high specificity is mandatory for avoiding cross-reaction in epidemiological contexts where the two species of Leishmania occur in sympatry.

Snakes and Souks: Zoonotic pathogens associated to reptiles in the Marrakech markets, Morocco.

The world-famous markets of Marrakech, also known in Arabic as souks, harbor a vast diversity of reptiles that are sold for medicinal/magic/pet purposes or used for snake charming. This unique epidemiological context has never been studied considering the interactions of humans, reptiles, and zoonotic pathogens. Thus, the aim of this study was to identify the parasites and pathogens present in blood and feces associated with handled reptiles in the markets of Marrakech to assess the risk of zoonotic transmission within the reptile-human interface. Privately owned reptiles (n = 118), coming from vendors or snake charmers, were examined and blood and feces sampled. DNA was extracted and molecular screening (cPCR, nPCR, qPCR, dqPCR) was performed aiming to identify potentially zoonotic pathogens (i.e., Anaplasma/Ehrlichia spp., Rickettsia spp., Borrelia burgdorferi sensu lato, Coxiella burnetii, Babesia/Theileria spp., Cryptosporidium spp., Giardia spp., Leishmania spp., Cestoda). Overall, 28.9% (34/118) of reptiles were positive for at least one pathogen. In blood, Anaplasma spp. were detected in four snakes, with two Montpellier snakes positive for Anaplasma phagocytophilum, while Rickettsia spp. were detected in one Mediterranean chameleon and four puff adders. Leishmania tarentolae was molecularly detected in a Mediterranean chameleon and a Montpellier snake. In feces, the cox1 gene generated a myriad of sequences for nematodes, cestodes, fungi and bacteria. Importantly, Proteus vulgaris was identified from a Mediterranean chameleon. Cryptosporidium spp. nPCR yielded a positive sample (i.e., Cryptosporidium sp. apodemus genotype I) from a Moroccan worm lizard, as well as for bacteria such as Pseudomonas aeruginosa in an Egyptian cobra, and Morganella morganii from a puff adder. Results from this study demonstrated the risk of zoonotic transmission of microorganisms and parasites present in blood and feces from reptiles that are brought to the souks in Marrakech, Morocco, to be sold for medicinal purposes or used for snake charming, being in direct and straight contact with humans.

Interaction between Wolbachia pipientis and Leishmania infantum in heartworm infected dogs.

BACKGROUND: Wolbachia is a Gram-negative endosymbiont associated with several species of arthropods and filarioid nematodes, including Dirofilaria immitis. This endosymbiont may elicit a Th1 response, which is a component of the immunity against Leishmania infantum. METHODS: To investigate the interactions between Wolbachia of D. immitis and L. infantum in naturally infected dogs and cytokine circulation, dogs without clinical signs (n = 187) were selected. Dogs were tested for microfilariae (mfs) by Knott, for female antigens of D. immitis by SNAP, and for anti-L. infantum antibodies by IFAT and assigned to four groups. Dogs of group 1 (G1) and 2 (G2) were positive for D. immitis and positive or negative to L. infantum, respectively. Dogs of group 3 (G3) and 4 (G4) were negative to D. immitis and positive or negative to L. infantum, respectively. Wolbachia and L. infantum DNA was quantified by real-time PCR (qPCR) in dog blood samples. A subset of dogs (n = 65) was examined to assess pro- and anti-inflammatory cytokine production using an ELISA test. RESULTS: Of 93 dogs positive to D. immitis with circulating mfs, 85% were positive to Wolbachia, with the highest amount of DNA detected in G1 and the lowest in dogs with low mfs load in G1 and G2. Among dogs positive to L. infantum, 66% from G1 showed low antibody titer, while 48.9% from G3 had the highest antibody titer. Of 37 dogs positive to Wolbachia from G1, 26 (70.3%) had low antibody titers to L. infantum (1:160). Among cytokines, TNFα showed the highest mean concentration in G1 (246.5 pg/ml), IFNγ being the one most represented (64.3%). IL-10 (1809.5 pg/ml) and IL-6 (123.5 pg/ml) showed the highest mean concentration in dogs from G1. A lower percentage of dogs producing IL-4 was observed in all groups examined, with the highest mean concentration (2794 pg/ml) recorded in G2. CONCLUSION: Results show the association of D. immitis and Wolbachia with the lower antibody titers of L. infantum in co-infected dogs, suggesting the hypothesis that the endosymbiont may affect the development of the patent leishmaniosis. However, due to the limitations associated with the heterogeneity of naturally infected dogs in field conditions, results should be validated by investigation on experimental models.

Assessment of circulating immune complexes in canine leishmaniosis and dirofilariosis.

Canine leishmaniosis (CanL) by Leishmania infantum (L.i.) and heartworm disease by Dirofilaria immitis (D.i.) are common zoonotic vector-borne diseases (VBDs) characterized by a variety of pathological and clinical signs. The immunopathology in both VBDs is extremely complex, and their clinical manifestations are strongly dependent on the type of immune response elicited by the parasites. In particular, the formation of circulating immune complexes (CICs) plays an important role in the pathogenesis of these VBDs. Based on the international guidelines, dogs with high anti-L. infantum antibody titres and one or more clinical and/or laboratory signs related to CanL require anti-Leishmania treatment. Consequently, the CICs measurement could be used for improving the clinical staging process of CanL. The aim of the study was to assess the CICs level by a competitive inhibition enzyme immunoassay, in healthy or sick dogs seropositive to L.i. and in healthy dogs positive to D.i.. Out of 51 enrolled dogs, 11 were included in Group A (seronegative to L.i., D.i. negative and healthy), 15 in Group B (exposed to L.i., D.i. negative and healthy), 12 in Group C (seropositive to L.i., D.i. negative and sick) and 13 in Group D (seronegative to L.i, D.i. positive and healthy). The comparison of CIC level in canine sera revealed a significant difference among groups (P < 0.001), with the highest concentration (i.e., median = 104.6 µg/mL) in dogs with CanL. The findings of the study highlight the CICs measurement as a useful tool in the clinical staging of CanL for avoiding misclassification of dogs as leishmaniotic, thus not requiring anti-Leishmania therapy, as well as the possibility of results misuse in geographical areas where both leishmaniosis and heart-worm disease are endemic.

Neurogenic Bladder in Dogs, Cats and Humans: A Comparative Review of Neurological Diseases.

Lower urinary tract disease (LUTD) includes abnormalities in the structure and function of the bladder and the urethra. LUTD caused by neurological disease is defined neurogenic bladder (NB). The integrity of the central nervous system (CNS) and peripheral nervous system (PNS) is required to explicate normal micturition, maintaining the proper function of bladder and urethra. The location and type of neurological lesions influence the pattern of clinical manifestations, potential treatment, and prognosis. Though, in dogs and cats, spinal cord injury is considered mainly responsible for bladder and/or urethra incompetence, other disorders, congenital or acquired, involving CNS or PNS, could play a role in NB. In veterinary medicine, the information about the epidemiology, prevalence, etiopathogenesis, diagnosis and treatment of NB are scattered. The aim of this study is to provide an overview of the epidemiology, prevalence, clinical findings, diagnosis and prognosis for NB in dogs and cats compared with humans.

Efficacy of domperidone plus renal diet in slowing the progression of chronic kidney disease in dogs with leishmaniosis.

BACKGROUND: Chronic kidney disease (CKD) represents the main cause of mortality in dogs with leishmaniosis. Domperidone has recently been reported to improve kidney function in leishmaniotic dogs affected by CKD. Serum symmetric dimethylarginine (sSDMA) has also been shown to be a useful biomarker for earlier detection of decreased kidney function when compared to serum creatinine (sCr). This study aimed to assess the efficacy of domperidone plus renal diet in slowing the progression of nephropathy in leishmaniotic dogs with CKD, evaluating sSDMA and sCr as markers of kidney function. METHODS: This study was a therapeutic, prospective, randomized, controlled, 11-month-long field trial. Dogs were recruited if classified as "exposed" to or "infected" with Leishmania infantum and affected by CKD at early stages. After enrolment (T0), dogs were randomized into groups T (treatment) and C (control). All dogs were fed a renal diet and then followed up at 90 (T1), 210 (T2), and 330 (T3) days after inclusion in the study. At T1 and T2, dogs in group T received an oral suspension of domperidone (1 ml/10 kg once a day for up to 28 days). RESULTS: Twenty-two dogs (i.e., n = 12 in group T and n = 10 in group C) completed the study. At T0, the entire population of enrolled dogs presented a mean sSDMA value of 16.5 ± 3.4 µg/dl. At T1 (i.e., after 3 months of renal diet), sSDMA was significantly decreased in both groups, with an sSDMA of 13.1 ± 4.4 µg/dl for the entire population involved. From T1 to T3, sSDMA gradually increased in group C, while remaining stable in group T, which continued to show a significantly lower value of sSDMA at T3 than at T0. Regarding sCr, at T0 and T1, the mean values of the entire population of dogs were 1.1 ± 0.3 and 1.0 ± 0.4 mg/dl, respectively, with no statistical differences between groups T and C. In group T, sCr decreased significantly from T0 to T1, while returning at T3 to values similar to T0. CONCLUSIONS: In this study, domperidone plus renal diet reduced the progression of kidney disease in leishmaniotic dogs affected by CKD.

Role of Gut Microbiota through Gut-Brain Axis in Epileptogenesis: A Systematic Review of Human and Veterinary Medicine.

Canine idiopathic epilepsy is a common neurological disease characterized by the enduring predisposition of the cerebral cortex to generate seizures. An etiological explanation has not been fully identified in humans and dogs, and, among the presumed causes, several studies support the possible involvement of gut microbiota. In this review, the authors summarize the evidence of the reasonable role of gut microbiota in epilepsy through the so-called gut-brain axis. The authors provide an overview of recent clinical and preclinical studies in humans and dogs in which the modulation of intestinal permeability, the alteration of local immune response, and the alteration in production of essential metabolites and neurotransmitters associated with dysbiosis could be responsible for the pathogenesis of canine epilepsy. A systematic review of the literature, following the PRISMA guidelines, was performed in two databases (PubMed and Web of Science). Eleven studies were included and reviewed supporting the connection between gut microbiota and epilepsy via the gut-brain axis.

Erythrocyte sedimentation rate in canine leishmaniosis diagnosis: A new resource.

This study aims to evaluate the erythrocyte sedimentation rate (ESR) in Leishmania infantum-seropositive dogs compared with healthy dogs and to assess the existence of a correlation between ESR and clinical form of Canine leishmaniosis (CanL) as well as acute phase proteins (APPs). From October 2021 to January 2022, dogs were recruited in this study if L. infantum-seropositive by enzyme-linked immunoassay and classified as exposed or affected by a CanL active form based on physical examination, clinical score, and laboratory results [i.e., complete blood count, biochemical panel such as C-reactive protein (CRP) and serum ferritin, serum protein electrophoresis, and fibrinogen concentration measurement]. To evaluate the ESR of the dogs, a point-of-care device was used with a reference interval of 0-10 mm/h. Moreover, the ESR evaluation has been also performed in clinically healthy dogs, as control group. Thirty-six L. infantum-seropositive dogs [i.e., exposed (n = 10) and affected by CanL active form (n = 26)] were included in the study. Twenty-two healthy dogs were also enrolled. The mean value of ESR in dogs affected by a CanL active form was significantly higher than in exposed and healthy dogs (p < 0.0001). The ESR level was increased in 92% of dogs with CanL active form while positive APPs such as CRP, fibrinogen, and serum ferritin were increased only in 46, 48, and 58% of the animals, respectively. In exposed dogs, the ESR level was increased in 40% of cases. In dogs with active form, a significant positive correlation between ESR and total proteins, globulins, CRP, and fibrinogen, as well as a significant negative correlation between ESR and hematocrit, hemoglobin, and albumin/globulin ratio were detected. This study provides for the first-time data on ESR in L. infantum-seropositive dogs demonstrating dogs affected by a CanL active form have the highest ESR level and the majority of these dogs presented an increased ESR compared with exposed and healthy dogs. The evaluation of ESR by a point-of-care device proved to be a simple, inexpensive, and ready-to-use benchtop tool and ESR can be considered a helpful and timely inflammatory biomarker for the diagnosis of a CanL active form.

Leishmania (Sauroleishmania) tarentolae isolation and sympatric occurrence with Leishmania (Leishmania) infantum in geckoes, dogs and sand flies.

The trypanosomatid protist Leishmania tarentolae is a saurian-associated parasite vectored by the Sergentomyia minuta sand fly. This study aimed to confirm the circulation of L. infantum and L. tarentolae in sand flies, reptiles and dogs and to isolate new strains of these protists. Reptilian and sheltered dog blood samples were collected, and sand flies were captured. Samples were tested for Leishmania spp. using duplex real-time PCR (dqPCR) and real-time PCR (qPCR); the origin of blood meal was identified in engorged sand flies by conventional PCR. The reptilian blood and intestinal content of sand fly females were cultured. Dog sera were tested by IFAT using both Leishmania species. Four Tarentola mauritanica geckoes were molecularly positive for L. infantum or L. tarentolae, with no co-infections; moreover, amastigote-like forms of L. infantum were observed in the bone marrow. 24/294 sand flies scored positive for Leishmania spp. by dqPCR, 21 S. minuta and two Phlebotomus perniciosus were positive for L. tarentolae, while only a single Ph. perniciosus was positive for L. infantum. Blood meal analysis confirmed reptile and dog in S. minuta, dog and human in Ph. perniciosus and dog in Phlebotomus neglectus. Two axenic strains of L. tarentolae were obtained. Twelve of 19 dogs scored positive for L. infantum and L. tarentolae by IFAT and three of them also for L. infantum by dqPCR, and six by qPCR. These data confirm the sympatric circulation of L. infantum and L. tarentolae in geckoes, sand flies, and dogs, and suggest that geckoes may be infected with L. infantum.

A review of the neural control of micturition in dogs and cats: neuroanatomy, neurophysiology and neuroplasticity.

This article discusses the current knowledge on the role of the neurological structures, especially the cerebellum and the hypothalamus, and compares the information with human medicine. Micturition is a complex voluntary and involuntarily mechanism. Its physiological completion strictly depends on the hierarchical organisation of the central nervous system pathways in the peripheral nervous system. Although the role of the peripheral nervous system and subcortical areas, such as brainstem centres, are well established in veterinary medicine, the role of the cerebellum and hypothalamus have been poorly investigated and understood. Lower urinary tract dysfunction is often associated with neurological diseases that cause neurogenic bladder (NB). The neuroplasticity of the nervous system in the developmental changes of the mechanism of micturition during the prenatal and postnatal periods is also analysed.

Prevalence of bacteriuria in cats with neurogenic bladder.

Urinary tract infections are defined as the adherence, multiplication, and persistence of an infectious agent within the urogenital system, causing an associated inflammatory response and clinical signs; instead, the presence of bacteria in urine as determined by positive bacterial culture (PUC) from a properly collected urine specimen, in the absence of clinical signs, is defined subclinical bacteriuria. Limited information on the prevalence of PUC in spinal cord injury cats affected by neurogenic bladder (NB) is available. On contrary, in NB dogs and humans the prevalence of bacteriuria is well documented. Moreover, while in humans information about bacteriemia associated with NB is already available, this aspect has never been studied in NB cats. The aim of this prospective study was to determine the prevalence of PUC in cats with NB, compared to animals affected by chronic kidney disease (CKD) and healthy cats. Furthermore, the prevalence of bacteriemia in cats with NB was evaluated. Fifty-one cats met the inclusion criteria: 12 cats were affected by NB, 22 had CKD and 17 were healthy. The prevalence of PUC was 58.33% and 18% in NB and CKD cat populations, respectively. All blood cultures were negative. The incomplete bladder emptying and the decreased resistance in the bladder wall could be considered predisposing elements to PUC in the NB feline population. The results of this study highlight, for the first time, an high prevalence of PUC in cats affected by NB, which was not found to be associated with bacteriemia.

Serum protein electrophoresis in Dirofilaria immitis naturally infected dogs: Latest news and a systematic literature review.

According to the main Guidelines on canine heartworm disease (HWD) by the American and European Societies (i.e., AHS, ESDA, and ESCCAP), a correct diagnosis of Dirofilaria immitis infection should include the detection of circulating microfilariae in the whole blood and the adult antigens in serum or plasma sample. So far, scant data are available on laboratory abnormalities in dogs affected by HWD, although techniques including serum protein electrophoresis (SPEP) have proved to be useful for the diagnosis and monitoring of other vector-borne diseases, such as the canine leishmaniosis. Therefore, this study aims to evaluate the SPEP pattern in dogs naturally infected by D. immitis. Furthermore, a systematic review of the literature on this topic was carried out. Medical records from heartworm-positive dogs, of any sex, age, and breed and with available clinical examination and laboratory test results (i.e., complete blood count, serum biochemical profile, and SPEP) were retrospectively collected. If available, laboratory results obtained from dogs after treatment for HWD were also evaluated. When compared with the reference intervals, out of 30 dogs infected by D. immitis and enrolled, 63.3% (n = 19) had a lower percentage of albumin, and 80.0% (n = 24) had higher percentages of beta globulins, with beta-2, and especially beta-3 globulins the most frequently altered fractions. In terms of absolute values (g/dL), the proportion of dogs with hypoalbuminemia, and increased total globulin, alpha, beta- and gamma globulins were 4/30 (13.3%), 6/30 (20.0%), 2/30 (6.7%), 16/30 (53.3%) and 8/30 (26.7%), respectively. For 7 dogs, SPEP results evaluated three and six months after treatment with doxycycline (10 mg/kg BID for 4 weeks) were available. In these dogs a significant post-treatment increase in the percentage of albumin, alpha-2 globulin, and albumin/globulins ratio was observed, as well as a significant decrease both in the percentage and in the absolute value of total-, beta-, and beta-3 globulins. The systematic review of literature databases yielded a total of three studies that were considered eligible and included in the qualitative synthesis. This study provides novel information on SPEP alterations in dogs naturally infected by D. immitis. The evaluation of serum proteins and their electrophoretic pattern may represent an important diagnostic tool for a prompt and accurate diagnosis (e.g., differentiating infections in dogs sharing similar clinical signs and endemic in the same geographical area) and monitoring of HWD.

The knowns and unknowns of the efficacy of neem oil (Azadirachta indica) used as a preventative measure against Leishmania sand fly vectors (Phlebotomus genus).

Since domestic dogs are the main reservoir hosts of Leishmania infantum throughout the world, they are the main focus in terms of controlling zoonotic visceral leishmaniosis. To protect dogs from leishmaniosis, chemical repellents of durable efficacy are available in the form of collars, spot-on and sprays. However, the negative effects of chemical pesticides on the environment are well established as they affect animals and plants. This phenomenon has created the need for safer and more environmentally friendly substitutes. Plant extract-based insecticides and/or repellents have therefore been increasingly used by pet owners and veterinarians. Several botanical products have been tested as insecticides and/or repellents against a variety of bloodsucking arthropods that transmit human diseases. Among the products tested against Leishmania vectors, neem oil containing azadirachtin is the most studied. This study reviews the scientific literature concerning the efficacy of neem oil (azadirachtin-based products) against phlebotomine sand fly bites. A questionnaire was also administered to assess Italian veterinarians' attitudes to the use of neem oil. The survey was anonymous and consisted of three closed-ended questions. According to the data reported in the literature, the efficacy of neem oil in reducing the risk of sand fly bites has been tested against Phlebotomus papatasi, Phlebotomus perniciosus, Phlebotomus argentipes, Phlebotomus orientalis and Phlebotomus bergeroti. The efficacy of the products tested was expressed in percentages, ranging from 74.9% to 100%. The protection time was only available for six out of eight studies, ranging from "only during the first hour" (minimum protection time) to "all night" (expected maximum protection time). As regards the attitude to recommending the use of neem oil, 208 veterinarians participated in the online survey. Of the 126 veterinarians who recommended natural products, 119 (94.44%) reported that they recommended the use of neem oil-based products. Considering the limited data on the duration of protection and the dose of the active ingredient, more studies are required on the efficacy of neem oil-based products in reducing the risk of contracting canine leishmaniosis. These studies should also refer specifically to the concentration of the active ingredient as well as the interval of administration. Until such results are available, the use of azadirachtin-based products as the only topical products for the prevention of leishmaniosis in dogs is not recommended.

Conjunctival Swab Real Time-PCR in Leishmania infantum Seropositive Dogs: Diagnostic and Prognostic Values.

Conjunctival swabs (CS) are the most promising non-invasive samples for the diagnosis and the regular screening of Leishmania infantum infection in dogs although knowledge on their diagnostic performance is still inconclusive. This study evaluates CS real time-PCR (qPCR) analysis for the diagnosis of canine leishmaniosis (CanL) and its prognostic value in seropositive dogs from an endemic area. In October 2020 (T0), 26 dogs were enrolled, divided in two groups according to anti-L. infantum antibody titres (n = 13, group low titre (LT) and n = 13, group high titre (HT)), and followed-up in August 2021. At both timepoints, animals underwent clinical examination, complete blood count and biochemical analyses, and serological (indirect fluorescent antibody test) and molecular (CS and peripheral blood qPCR) testing. At T0, 10 out of 26 enrolled dogs were positive at CS qPCR, with the number of positive animals significantly higher in group HT than in LT. After 10 months, only 5 out of 21 dogs that completed the trial still tested CS qPCR positive, and none of them developed an active CanL based on clinical score and antibody titre. None of the dogs required any leishmanicidal and/or leishmaniostatic treatments. This prospective study showed unsatisfying diagnostic and prognostic performances of CS qPCR analysis in L. infantum seropositive asymptomatic dogs from an endemic area.

Is proteinuria a rare condition in apparently healthy and sick cats? A feline practice experience (2007-2018).

Background: Proteinuria is assumed to be less frequent in cats than in dogs and is mainly associated with chronic kidney disease (CKD). Aim: The current study aimed to evaluate and compare urine protein-to-creatinine (UPC) values retrospectively in cats visited for comprehensive annual health check or for presenting systemic clinical signs related to CKD. Methods: UPC ratio was retrospectively evaluated in 112 owned cats, out of which 51 (45.5%) were apparently healthy cats according to their owners who visited for comprehensive annual health checks and 61 (54.5%) sick cats, presenting systemic clinical signs suggesting CKD, such as weight loss or polyuria/polydipsia, among others. Results: Based on UPC, the present study found that 54.5% of all cats included were borderline proteinuric or proteinuric, having increased UPC (UPC ≥ 0.2), with 35.7% included in the sick group and 18.7% in the health-check group. Increased UPC was also statistically associated with azotemia and isosthenuria (urinary-specific gravity between 1,008 and 1,035) in both sick and health-check groups of cats. Conclusion: Independent of the reason for their medical visit, it could be concluded that borderline proteinuria and proteinuria were statistically mainly related to CKD in cats. Furthermore, the measurement of UPC could be very useful in the detection and management of CKD in apparently healthy cats during a medical visit for annual health check irrespective of the age.

Effect of domperidone (leisguard®) on antibody titers, inflammatory markers and creatinine in dogs with leishmaniosis and chronic kidney disease.

BACKGROUND: Immunotherapeutic drugs, such as domperidone, have been shown to be promising treatments against canine leishmaniosis (CanL), but limited data are available. The aim of this pilot study (therapeutic, prospective and non-controlled) was to evaluate the effect of domperidone on serum antibody titers of Leishmania infantum, globulins, gamma globulins, acute-phase proteins (e.g. C-reactive protein [CRP]), big endothelin-1 (big ET-1), serum creatinine (SC) and proteinuria in dogs with leishmaniosis affected by chronic kidney disease (CKD). METHODS: Dogs were recruited if "exposed" to or "infected" with L. infantum and affected by CKD (IRIS stage 1 [proteinuric] or IRIS stage 2-3a [SC < 3.5 mg/dl; proteinuric or non-proteinuric]). After inclusion, an oral suspension of domperidone was administered, and the dogs were followed up for 180 days, with checks at 30, 60, 90 and 180 days after initial treatment. RESULTS: Of the 14 recruited dogs, nine showed a statistically significant reduction in SC (χ2 = 9.1, df = 3, P = 0.028), but not in the urine protein/creatinine ratio (χ2 = 6.43, df = 3, P = 0.092). All dogs showed a significant reduction in antibody titers for L. infantum (χ2 = 9.56, df = 2, P = 0.008), globulins (χ2 = 11.08, df = 3, P = 0.011) and gamma globulins (χ2 = 12.38, df = 3, P = 0.006) during the study period. There was also a statistically significant reduction in CRP (χ2 = 16.7, df = 3, P = 0.001), but not in big ET-1 (χ2 = 2.04, df = 3, P = 0.563). CONCLUSIONS: This study provides preliminary results on the ability of domperidone to improve SC and reduce anti-L. infantum antibody titers, globulins, gamma globulins and CRP in dogs with leishmaniosis and CKD.

Detection of Leishmania tarentolae in lizards, sand flies and dogs in southern Italy, where Leishmania infantum is endemic: hindrances and opportunities.

BACKGROUND: Leishmania tarentolae is a protozoan isolated from geckoes (Tarentola annularis, Tarentola mauritanica), which is considered non-pathogenic and is transmitted by herpetophilic Sergentomyia spp. sand flies. This species occurs in sympatry with Leishmania infantum in areas where canine leishmaniasis is endemic. In the present study, we investigated the circulation of L. tarentolae and L. infantum in sand flies, dogs and lizards in a dog shelter in southern Italy, where canine leishmaniasis by L. infantum is endemic. METHODS: Sheltered dogs (n = 100) negative for Leishmania spp. (March 2020) were screened by immunofluorescence antibody test (IFAT) using promastigotes of both species at two time points (June 2020 and March 2021). Whole blood from dogs, tissues of Podarcis siculus lizards (n = 28) and sand flies (n = 2306) were also sampled and tested by a duplex real-time PCR (dqPCR). Host blood meal was assessed in sand flies by PCR. RESULTS: Overall, 16 dogs became positive for L. infantum and/or L. tarentolae by IFAT at one or both sampling periods. One canine blood sample was positive for L. infantum, whilst two for L. tarentolae by dqPCR. At the cytology of lizard blood, Leishmania spp. amastigote-like forms were detected in erythrocytes. Twenty-two tissue samples, mostly lung (21.4%), scored molecularly positive for L. tarentolae, corresponding to 10 lizards (i.e., 35.7%). Of the female Sergentomyia minuta sampled (n = 1252), 158 scored positive for L. tarentolae, four for L. infantum, and one co-infected. Two Phlebotomus perniciosus (out of 29 females) were positive for L. tarentolae. Engorged S. minuta (n = 10) fed on humans, and one P. perniciosus, positive for L. tarentolae, on lagomorphs. CONCLUSIONS: Dogs and lacertid lizards (Podarcis siculus) were herein found for the first time infected by L. tarentolae. The detection of both L. tarentolae and L. infantum in S. minuta and P. perniciosus suggests their sympatric circulation, with a potential overlap in vertebrate hosts. The interactions between L. tarentolae and L. infantum should be further investigated in both vectors and vertebrate hosts to understand the potential implications for the diagnosis and control of canine leishmaniasis in endemic areas.

Efficacy of afoxolaner (NexGard®) in preventing the transmission of Leishmania infantum and Dirofilaria immitis to sheltered dogs in a highly endemic area.

BACKGROUND: Leishmania infantum and Dirofilaria immitis are among the most important canine vector-borne pathogens (CVBPs) of zoonotic concern in Europe. In endemic areas for both of these CVBPs, the use of systemic ectoparasiticides, such as afoxolaner (NexGard®; Boehringer Ingelheim Animal Health), may have the potential for controlling these infections. The aim of this study was to assess, for the first time, the insecticidal efficacy of NexGard® in decreasing the transmission of D. immitis and L. infantum to sheltered dogs living in a hyperendemic area, compared to the year before treatment, as well as its impact on the abundance of mosquito and sand fly populations. METHODS: All dogs (n = 179) enrolled in the study were divided into two groups based on their infection status at enrollment: a non-infected group (G1) and an infected group (G2; infected with D. immitis, L. infantum or both). The study was conducted from March 2020 to March 2021. In order to exclude all animals infected with L. infantum and D. immitis before March 2020 (sampling time: T0), dogs in G1 were sampled in June (T1; i.e. T0 + 90 days) and in October 2020 (T2; i.e. T0 + 210 days). From March to September 2020, all animals (G1 and G2) were weighed and treated monthly with NexGard®. Animals in G1 were tested for the last time in March 2021 (T3; i.e. T0 + 330 days) for assessing post-treatment incidence rate of infection and prevention efficacy. RESULTS: The post-treatment incidence of D. immitis was 3.7% (1/27; 95% confidence interval [CI]: 0.2-18.1) and that of L. infantum was 3.6% (3/83; 95% CI: 1.0-10.1). Considering the annual incidence in 2019 and 2020, the protective efficacy against D. immitis and L. infantum infections was 94.2 and 64%, respectively. Of the female mosquitoes collected (n = 146), only one pool out of 50 tested positive for D. immitis DNA, whereas out of 1252 female Sergentomya minuta specimens collected, only four tested positive for L. infantum (0.3%). CONCLUSIONS: Afoxolaner is efficacious in decreasing the rate of transmission of both D. immitis and L. infantum; however, comparison of the pre- and post-treatment period demonstrated that there was a significant difference only in the seasonal incidences of D. immitis infection. Preventive measures are recommended throughout the year in endemic areas to reduce the risk of pathogen transmission to animals and humans.