Collaborating to Cure the Most Common Parasites on the Planet

Soil-transmitted helminths (STHs), most notably, hookworms, whipworms, and Ascaris, are nematodes that infect more than 1.5 billion of the poorest people and are leading causes of morbidity worldwide. Only one class of de-worming drugs (anthelmintic) is commonly used in mass drug administrations. New anthelmintics are urgently needed to overcome emerging resistance and to produce higher cure rates. Crystal (Cry) proteins, in particular Cry5B, made by Bacillus thuringiensis (Bt) are promising new candidates. Cry5B has excellent anthelmintic properties against many free-living and parasitic nematodes, including in vivo efficacy against multiple STH infections in rodents (Heligomasmidoes polygyrus and Ancylostoma ceylanicum) and in pigs (Ascaris suum). An enormous challenge for STHs, very different from most diseases worked on in the developing world, is the requirement that therapies be very cheap (the people infected are very poor and current drugs costs pennies a dose), massively scalable (over 4 billion people are at risk from infection), and have a long shelf life in harsh environments, that have high temperature and humidity and no cold chain. Working together, we have made excellent progress in our development efforts to produce a deployable version of Cry5B that is cheap, safe, scalable, and stable. These efforts are focused on microbiology, bacterial engineering, expression, and formulation. In the process of this work, we have discovered a novel bacterial expression system that meets these key requirements. In addition, we will provide latest information about the broad spectrum of activity of Cry5B against key parasites that make this therapeutic a very attractive alternative from current treatments

Anti-oxidant effect of N-acetyl cysteine in dogs with chronic kidney disease

The present study was carried out with the objectives of assessing oxidative stress in dogs with chronic kidney disease (CKD) and evaluating response to treatment with N-acetyl cysteine (NAC). Dogs diagnosed with stage III CKD as per the guidelines of the International Renal Interest Society (IRIS) were included in the study. The animals were divided into two groups. Animals of one group were given standard therapy for CKD and the animals of the second group were administered NAC along with standard therapy. Oxidative stress parameters such as total antioxidant status (TAS), serum malondialdehyde (MDA) level and plasma glutathione peroxidase (GSH-Px) activity were studied. On the day of presentation, a significant increase in the mean values of serum MDA and TAS were observed in diseased animals compared to healthy animals, whereas a significant decline was noted in plasma GSH-Px activity. After treatment, a significant decline in serum MDA and TAS were recorded in animals of group II receiving NAC therapy. A significant increase in plasma glutathione GSH-Px activity was recorded in this group. N-acetyl cysteine therapy was found to be effective in the management of oxidative stress in dogs with chronic kidney disease

A new paraprobiotic-based treatment for control of Haemonchus contortus in sheep

Haemonchus contortus is a critical parasite of goats and sheep. Infection by this blood-feeding gastrointestinal nematode (GIN) parasite has significant health consequences, especially in lambs and kids. The parasite has developed resistance to virtually all known classes of small molecule anthelmintics used to treat it, giving rise in some areas to multidrug resistant parasites that are very difficult to control. Thus, new anthelmintics are urgently needed. Bacillus thuringiensis (Bt) crystal protein 5B (Cry5B), a naturally occurring protein made by a bacterium widely and safely used around the world as a bioinsecticide, represents a new non-small molecule modality for treating GINs. Cry5B has demonstrated anthelmintic activities against parasites of monogastric animals, including some related to those that infect humans, but has not yet been studied in a ruminant. Here we show that H. contortus adults are susceptible to Cry5B protein in vitro. Cry5B produced in its natural form as a spore-crystal lysate against H. contortus infections in goats had no significant efficacy. However, a new Active Pharmaceutical Ingredient (API) paraprobiotic form of Cry5B called IBaCC (Inactivated Bacterium with Cytosolic Crystals), in which Cry5B crystals are encapsulated in dead Bt cell wall ghosts, showed excellent efficacy in vitro against larval stages of H. contortus and relative protein stability in bovine rumen fluid. When given to sheep experimentally infected with H. contortus as three 60 mg/kg doses, Cry5B IBaCC resulted in significant reductions in fecal egg counts (90%) and parasite burdens (72%), with a very high impact on female parasites (96% reduction). These data indicate that Cry5B IBaCC is a potent new treatment tool for small ruminants in the battle against H. contortus

Haemato-biochemical and trichographic studies on mucocutaneous lesions in dogs

The present study was carried out with the objective of assessment of haematobiochemical and trichographic features of mucocutaneous lesions in dogs. Dogs presented with mucocutaneous lesions associated with various dermatological conditions were included in the study. Mange, atopic dermatitis, dermatophytosis, juvenile cellulitis, pyoderma and hypothyroidism with secondary Malassezia dermatitis were presented with mucocutaneous lesions in the perioral, periorbital and in the nostrils with least involvement of anogenital region. Haemato-biochemical analysis revealed anaemia, non-significant increase in total leucocyte count, neutrophilia, eosinophilia, elevated platelet count, hyperproteinaemia and hypoalbuminaemia. Elevated serum C-reactive protein was evident in all cases. Trichographic features revealed that highest number of hairs were included in the telogen phase of hair cycle with greater number of primary hairs. Hair fractures were evident in 20.8 per cent of hairs and trichoptilosis of hair tip was observed in 50.8 per cent of hairs studied. Demodex mites could be detected by hair pluck examination in all cases of demodicosis

A retrospective study of acute kidney injury in dogs with renal resistive index as a prognostic indicator

Doppler ultrasonography is relatively a new method for evaluation of the renal diseases in dogs. This study was conducted to evaluate the efficacy of renal resistive index (RRI) in diagnosis and prognosis of acute kidney injury (AKI) in dogs and to ascertain its correlationwith other renal parameters. Twenty-seven dogs with AKI along with ten clinically healthy dogs were evaluated. The aetiology of AKI was multifactorial which included viper snake envenomation, pyometra, Babesiagibsoni infection, canine parvo viral infection, Ehrlichiacanis infection, leptospiral infection, bone meal poisoning and tetracycline-induced nephrotoxicity. Dogs with AKI were characterized into five gradesas per the guidelines provided by International Renal Interest Society.Parameters such asblood urea nitrogen (BUN), creatinine, urine output, urine protein-creatinine (UPC) ratio and blood pressure were estimated. Both BUN and creatinine was significantly higher in affected dogs. Majority of the dogs with AKI were oliguric and had the presence of proteinuria, occult blood and leucocytes on urine dipstick examination. Urine Protein-Creatinine ratio was increased whereas no significant variation was noticed in blood pressure of dogs with AKI.B-mode ultrasonography of kidneys revealedrenomegaly, hyperechoic cortex, decreased cortico-medullary differentiation and loss of architecture in majority of the dogs with AKI. Renal resistive index was significantly increased inAKI and a positive correlation was established between RRI and serumcreatinine but not with UPC ratio, blood pressure and size of kidneys. Re-evaluation of RRIseven days post treatment showed considerablereduction. It wasconcludedthat an increase in RRIwas associated with severity of renal parenchymal injuryin AKI and reduction of RRI post-treatment indicated favorable prognosis

Comparative evaluation of darbepoetin therapy in non-regenerative anaemia associated with Babesia gibsoni infection in dogs

The present study describes a comparative evaluation of haemato-therapeutic response to darbepoetin therapy in non-regenerative anaemia associated with Babesia gibsoni infection in dogs. A total of 68 dogs, presented at Teaching Veterinary Clinical Complex, Mannuthy with signs of anaemia, weakness, anorexia and pallor of mucous membranes were screened for nonregenerative anaemia. Twenty dogs with a reticulocyte count of < 60,000/μL, haematocrit of < 30 per cent and positive for B. gibsoni were selected for detailed study. The twenty dogs were divided into two groups of ten animals and were subjected to estimation of complete blood count and analysis of serum protein and iron status. Ten dogs brought to the hospital for vaccination or health check-up served as healthy control. Haematological examination revealed significant anaemia and thrombocytopenia in both the groups. Serum total protein, albumin and total iron binding capacity revealed no difference between diseased groups with the healthy control. Serum iron and percentage transferrin saturation were similar in Group I and II, but significantly higher than healthy control. To compare darbepoetin efficacy, Group I dogs were administered with clindamycin and doxycycline for three weeks, whereas Group II was given with darbepoetin at the dose rate of 0.5 μg/kg body weight at weekly intervals for three weeks in addition to clindamycin-doxycycline as in Group I. Dogs of both groups showed complete remission of clinical signs within 10 days of therapy. Statistical analysis revealed no significant difference in haematological parameters between the treatment groups. It can be concluded from this study that there is no added advantage of darbepoetin therapy over conventional therapies against non-regenerative anaemia associated with B. gibsoni infection in dogs

Subclinical ketosis: Prevalence and risk factor analysis in early lactation dairy cattle of Thrissur district

Present study was conducted in Kerala Veterinary and Animal Sciences University farms, various organized and unorganised farms and individual households in Thrissur district during the period of May 2021 to April 2022 with the objectives of estimation of prevalence of subclinical ketosis (SCK) by screening dairy cows on 14th and 28th day post-partum. Multiparous cows with a blood beta-hydroxy butyrate (BHB) value ≥ 1.2 mmol/L without any overt clinical signs were considered as positive for SCK. One hundred and fourteen animals from second to seventh lactation were screened for SCK. Prevalence of SCK was 23.7 per cent on 14th day and 14.04 per cent on 28th day with a total prevalence of 27.2 per cent. Occurrence of SCK was highest in animals in second lactation (54.8 per cent) with more occurrence in crossbred Holstein Friesian cows. Maximum number of positive cases were observed in dairy cows of age between four to six years (54.8 per cent). Body condition score of diseased animals were higher than normal animals. Prevalence was highest in animals calved between months of July to September and 58.1 per cent of affected animals had male calf. Concurrent abnormalities during transition period were also assessed based on clinical examination and farm records

B-mode and Doppler ultrasound features of mammary neoplasms and their comparison with normal mammary glands in dogs

Canine mammary neoplasms are naturally occurring non-homogenous group of tumours with many resemblances to human breast cancer. In female dogs mammary tumours are of great clinical relevance due to the high prevalence and mortality rate, which varies according to the histopathological classification and clinical stage. The aim of the study was to compare the ultrasonographic features of the normal mammary gland with benign and malignant mammary neoplasms in dogs, through assessed by B-mode and Doppler mode ultrasonography. Ultrasonographic examination of seven normal mammary glands along with six benign and 12 malignant mammary neoplasms were performed. Among the parameters evaluated by B-Mode ultrasonography, significant differences were found (p < 0.01) in the tumour margin and invasiveness of neoplasms, where malignant tumours were invasive with uncircumscribed margin. Significant difference was also found in the presence of posterior acoustic enhancement (p<0.05) between benign and malignant mammary neoplasms. Elevated peak systolic velocity of blood flow within the tumour vessels in malignant mammary neoplasm was the only feature assessed with Doppler mode having significant difference

A New Paraprobiotic-based Treatment for Control of Haemonchus Contortus in Sheep

Haemonchus contortus is a critical parasite of goats and sheep. Infection by this blood-feeding gastrointestinal nematode (GIN) parasite has significant health consequences, especially in lambs and kids. The parasite has developed resistance to virtually all known classes of small molecule anthelmintics used to treat it, giving rise in some areas to multidrug resistant parasites that are very difficult to control. Thus, new anthelmintics are urgently needed. Bacillus thuringiensis (Bt) crystal protein 5B (Cry5B), a naturally occurring protein made by a bacterium widely and safely used around the world as a bioinsecticide, represents a new non-small molecule modality for treating GINs. Cry5B has demonstrated anthelmintic activities against parasites of monogastric animals, including some related to those that infect humans, but has not yet been studied in a ruminant. Here we show that H. contortus adults are susceptible to Cry5B protein in vitro. Cry5B produced in its natural form as a spore-crystal lysate against H. contortus infections in goats had no significant efficacy. However, a new Active Pharmaceutical Ingredient (API) paraprobiotic form of Cry5B called IBaCC (Inactivated Bacterium with Cytosolic Crystals), in which Cry5B crystals are encapsulated in dead Bt cell wall ghosts, showed excellent efficacy in vitro against larval stages of H. contortus and relative protein stability in bovine rumen fluid. When given to sheep experimentally infected with H. contortus as three 60 mg/kg doses, Cry5B IBaCC resulted in significant reductions in fecal egg counts (90%) and parasite burdens (72%), with a very high impact on female parasites (96% reduction). These data indicate that Cry5B IBaCC is a potent new treatment tool for small ruminants in the battle against H. contortus

LncRNA VEAL2 regulates PRKCB2 to modulate endothelial permeability in diabetic retinopathy

Long non‐coding RNAs (lncRNAs) are emerging as key regulators of endothelial cell function. Here, we investigated the role of a novel vascular endothelial‐associated lncRNA (VEAL2) in regulating endothelial permeability. Precise editing of veal2 loci in zebrafish (veal2 (gib005Δ8/+)) induced cranial hemorrhage. In vitro and in vivo studies revealed that veal2 competes with diacylglycerol for interaction with protein kinase C beta‐b (Prkcbb) and regulates its kinase activity. Using PRKCB2 as bait, we identified functional ortholog of veal2 in humans from HUVECs and named it as VEAL2. Overexpression and knockdown of VEAL2 affected tubulogenesis and permeability in HUVECs. VEAL2 was differentially expressed in choroid tissue in eye and blood from patients with diabetic retinopathy, a disease where PRKCB2 is known to be hyperactivated. Further, VEAL2 could rescue the effects of PRKCB2‐mediated turnover of endothelial junctional proteins thus reducing hyperpermeability in hyperglycemic HUVEC model of diabetic retinopathy. Based on evidence from zebrafish and hyperglycemic HUVEC models and diabetic retinopathy patients, we report a hitherto unknown VEAL2 lncRNA‐mediated regulation of PRKCB2, for modulating junctional dynamics and maintenance of endothelial permeability