Magnesium fertilization reduces high-temperature damages during anthesis in spring wheat (Triticum aestivum L.) by affecting pollen viability and seed weight.

Terminal heat during reproductive stages of wheat (Triticum aestivum L.) limits the productivity of the crop. Magnesium (Mg) is an essential macronutrient that is involved in many physiological and biochemical processes to affect photosynthesis and seed weight. The present study comparatively evaluated Mg applied to soil (80 kg MgSO4·7H2O ha-1) and to plant foliage (4% w/v) in improving wheat performance under terminal heat. Wheat crop was grown in two sets of treatments until the booting stage, and then one set of plants was shifted to a glasshouse (±5 °C) at the booting stage to grow until maturity in comparison to control plants kept under ambient warehouse condition. Heat stress reduced the pollen viability while foliar- and soil-applied Mg improved it by 3% and 6% under heat stress, respectively, compared to the control without Mg treatment. The 100-seed weight, spike length, and biological yield reduced by 39%, 19%, and 50% under heat stress; however, foliar and soil application increased 100-seed weight by 45% and 40%, spike length by 8% and 5%, and biological yield by 35% and 25% under heat stress, respectively. Soil Mg showed maximum SPAD chlorophyll values; however, response was statistically similar to that of foliar Mg as compared to the control without Mg supply. Membrane stability decreased (4%) due to heat stress while foliar and soil treatments improved membrane stability by 8% and 5% compared to that of the control, respectively. Thus, Mg application through soil or plant foliage can be an effective way to reduce negative impacts of terminal heat in wheat by improving pollen viability at anthesis and 100-seed weight that was attributed to increased chlorophyll contents during anthesis.

Cardio-Oncology: Managing Cardiovascular Complications of Cancer Therapies.

This narrative review explores the complex relationship between cancer medicines and cardiovascular health in the junction of oncology and cardiology, known as cardio-oncology. The study examines the historical development of cancer treatments and highlights the growing importance of cardiovascular problems in patient care. This text delves into the topic of cardiotoxicity, examining both conventional chemotherapeutic drugs like anthracyclines and more recent tyrosine kinase and immune checkpoint inhibitors. The complex molecular and cellular mechanisms that control cardiovascular problems are explained, including an understanding of how genetic predisposition influences an individual's sensitivity. The narrative expands into the crucial realm of risk stratification and evaluation, revealing advanced instruments for identifying cardiovascular risk in cancer patients. The importance of non-invasive imaging methods and biomarkers in early detection and continuous monitoring is emphasized. The prioritization of preventive tactics emphasizes the need to take proactive measures incorporating therapies to protect the heart throughout cancer treatment. It also highlights the significance of making lifestyle improvements to reduce risk factors. The narrative emphasizes the changing collaborative treatment environment, advocating for merging oncologists and cardiologists in a coordinated endeavor to maximize patient outcomes. In addition to clinical factors, the review explores the critical domain of patient education and support, acknowledging its crucial role in promoting informed decision-making and improving overall patient well-being. The latter portions of the text anticipate and consider upcoming treatments and existing research efforts that offer the potential for the future of cardio-oncology. This review seeks to provide a detailed viewpoint on the intricate connection between cancer treatments and cardiovascular well-being. Its objective is to encourage a more profound comprehension of the subject and prompt careful contemplation regarding the comprehensive care of cancer patients who confront the intricate difficulties presented by their treatment plans.

Evaluation of anti-inflammatory and analgesic effects of Hertia intermedia (Boiss.) Kuntze extract.

Hertia intermedia is a traditional medicinal plant of Balochistan, used for pain management and stomach problems. Current research work was intended to evaluate the anti-inflammatory and analgesic activities of crude ethanolic extract of H. intermedia. Anti-inflammatory activity was determined by the carrageenan-induced and histamine-induce Rat paw edema in rats, analgesic activity was determined by acetic acid-Induced writhing test, formalin-induced hind paw licking in mice and Tail immersion test. H. intermedia crude ethanolic extract showed significant (p<0.05) effect in both carrageenan and histamine-induced rat paw edema at both 250 and 500 mg/kg oral doses. There were significant analgesic activities in comparison with standard drug and control (p<0.05). It is concluded that H. intermedia crude ethanolic extract possesses significant anti-inflammatory and analgesic effects. However further studies may be carried out to isolate the phytochemicals responsible for anti-inflammatory and analgesic activities.

Measurement of age-dependent radiation ingestion doses due to gross alpha and gross beta exposure from medicinal plants.

This article presents the results of a study conducted to measure the gross alpha, gross beta activities in medicinal plant samples collected from different districts of Azad Kashmir, Pakistan. The ASC-950-DP gasless high-speed counter was used for the measurement of gross α/ß activities. Measured activities have been used to assess age-dependent annual effective doses for infants, one-, five-, ten-, and fifteen-year-old and adult people. For a medicinal plant consumption rate (MPCR) of 1.8 kg a-1, the average gross alpha and beta annual committed effective dose (ACED) delivered to one-, five-, ten-, fifteen-year-olds and adults fall below the WHO recommended level (290 µSv a-1) and that reported in the UNSCEAR 2000 (0.3 mSv a-1) report. Results obtained for the current study show that the radiological hazard related to the consumption of natural radionuclides in medicinal plants is inconsequential with exception of the ACED delivered to infants at an MPCR of 1.8 g a-1 and higher values.

Effects of Ferula asafetida, closantel, albendazole, oxfendazole, and ivermectin against Haemonchus contortus in goats and sheep.

BACKGROUND: Haemonchosis is a fatal disease of small ruminants caused by the parasite Haemonchus contortus (H. contortus). The most common drugs used in the treatment of H. contortus include albendazole, oxfendazole, and ivermectin. However, as previously reported in the treatment of haemonchosis, these medicines have acquired drug resistance problems over time. Interestingly, natural plant compounds have demonstrated promising effects in the treatment of H. contortus. Therefore, the current study evaluated the effects of plant extract, Ferula asafetida, against common drugs such as albendazole, oxfendazole, ivermectin, and closantel for the treatment of haemonchosis in small ruminants. METHODOLOGY: The current study was conducted on different small ruminant farms in Kasur District, Punjab, Pakistan. The positive animals (n = 720) after coprological examination were selected in this study and divided into two major groups (n = 360 goats and n = 360 sheep). Further, animals were divided into five treatment groups (A-E) and one control group with no treatment (F). Albendazole, oxfendazole, ivermectin, closantel, and Ferula asafetida were administered orally to groups A-E, respectively. The eggs per gram feces (EPG) were determined through the McMaster technique on days 0, 7th, and 14th of treatment. RESULTS: The results showed a significantly higher efficacy of closantel and Ferula asafetida against H. contortus in both goats (100% and 70%; p < 0.05) and sheep (99% and 87%; p < 0.05), respectively. No correlation was observed between EPG reduction with age and gender in both goats and sheep. CONCLUSION: Allopathic drug closantel and herbal drug, Ferula asafetida, have been proved an effective dewormer against H. contortus in small ruminants.

Evaluation of cardio protective, anti-inflammatory, analgesic and CNS depressant activity of Grewia asiatica L. fruit extract.

Grewia asiatica L. is a potential medicinal plant used for various diseases in traditional medicine. Current study was aimed to evaluate the cardio protective, anti-inflammatory, analgesic and CNS depressant activities of Grewia asiatica L. fruit extract. In cardio protective activity myocardial injury was produced by injection of Isoproterenol (200 mg/kg, s.c), G. asiatica 250 and 500mg/kg treated groups significantly (p<0.05) decreased the level of serum AST, ALT, LDH and CKMB, hence produced cardio protective effect. In analgesic activities G. asiatica produced significant (p<0.05) analgesic effects in acetic acid induced writhing, formalin, paw pressure and tail immersion test. G. asiatica at 250 and 500mg/kg oral dose, significantly (p<0.05) reduced the rat paw edema in carrageen an induced rat paw edema test. G. asiatica extract also produced significant CNS depressant effects in open field, hole board and thiopental sodium induced sleeping time. Findings of the current study suggest that G. asiatica fruit extract showed potential pharmacological effects and can be utilized in alternative medicine.

Practical High-Throughput Method to Screen Compounds for Anthelmintic Activity against Caenorhabditis elegans.

In the present study, we established a practical and cost-effective high throughput screening assay, which relies on the measurement of the motility of Caenorhabditis elegans by infrared light-interference. Using this assay, we screened 14,400 small molecules from the "HitFinder" library (Maybridge), achieving a hit rate of 0.3%. We identified small molecules that reproducibly inhibited the motility of C. elegans (young adults) and assessed dose relationships for a subset of compounds. Future work will critically evaluate the potential of some of these hits as candidates for subsequent optimisation or repurposing as nematocides or nematostats. This high throughput screening assay has the advantage over many previous assays in that it is cost- and time-effective to carry out and achieves a markedly higher throughput (~10,000 compounds per week); therefore, it is suited to the screening of libraries of tens to hundreds of thousands of compounds for subsequent evaluation and development. The present phenotypic whole-worm assay should be readily adaptable to a range of socioeconomically important parasitic nematodes of humans and animals, depending on their dimensions and motility characteristics in vitro, for the discovery of new anthelmintic candidates. This focus is particularly important, given the widespread problems associated with drug resistance in many parasitic worms of livestock animals globally.

Use of Medicinal Plants for Respiratory Diseases in Bahawalpur, Pakistan.

The most common ethnomedicinal plants being effective in respiratory disorders were studied for the first time in Bahawalpur District. The herbal medication represents a low-cost treatment for the local community. There is a need for documenting the traditional uses of plants for further investigation of bioactive compounds. Using a qualitative approach, the ethnobotanical data was collected from the district of Bahawalpur, Pakistan, from February 2018 to February 2020 through semistructured interviews with the local people and traditional healers. The quantitative analysis included use value, informant consensus factor, family importance value, and relative frequency citation. A total of 20 indigenous plants belonging to 17 families were documented from 185 informants. These plants were claimed to be used for the treatment of 10 respiratory ailments. The plant habit, part of the plant used, and mode of preparation were standardized for authentication. The herbs are the most used life form (55%), while trees and shrubs are also used. Leaves dominate with high use value (47.62%) followed by fruit, stem, flower, and other parts of plants. For the preparation of traditional remedies, decoction (76.19%) and extract (71.43%) are common preparation methods. However, other methods of paste infusion, powder juice, and ash are used to a lower extent. The plants with higher use value are Glycyrrhiza glabra, Acacia arabica, and Mentha piperita; these have significant potential therapeutic activity for respiratory disease. The ethnomedicinal importance of plants against respiratory diseases used by the local population (traditional healers) is the commercial availability of the herbal product. It is a first-time study in this area to fill the gap between traditional practices and synthetic medicine to screen out the phytochemical and pharmacological properties of plants that have a highly futuristic use value to develop antibiotic drug with least side effects by using sustainable methods.

1-Methyl-1H-pyrazole-5-carboxamide Derivatives Exhibit Unexpected Acute Mammalian Toxicity.

A series of 1-methyl-1H-pyrazole-5-carboxamides were synthesized as potent inhibitors of the parasitic nematode of sheep, Haemonchus contortus. These compounds did not show overt cytotoxicity to a range of mammalian cell lines under standard in vitro culture conditions, had high selectivity indices, and were progressed to an acute toxicity study in a rodent model. Strikingly, acute toxicity was observed in mice. Experiments measuring cellular respiration showed a dose-dependent inhibition of mitochondrial respiration. Under these conditions, potent cytotoxicity was observed for these compounds in rat hepatocytes suggesting that the potent acute mammalian toxicity of this chemotype is most likely associated with respiratory inhibition. In contrast, parasite toxicity was not correlated to acute toxicity or cytotoxicity in respiring cells. This paper highlights the importance of identifying an appropriate in vitro predictor of in vivo toxicity early on in the drug discovery pipeline, in particular assessment for in vitro mitochondrial toxicity.

Interactive effect of exogenous protease enzyme and dietary crude protein levels on growth and digestibility indices in broiler chickens during the starter phase.

This study was conducted to study the effect of protease as well as three dietary crude protein levels on growth and nutrient digestibility in broilers. The broiler chickens (n = 540) were assigned to six dietary treatments with six replicates. Three dietary crude protein (CP) levels (17, 19, and 21%) along with two levels of protease (0 and 30,000 IU/kg) were investigated in 3 × 2 factorial arrangement during the starter phase. Body weight was higher (P < 0.05) in group fed with CP-21; however, feed intake was significantly (P < 0.05) lower in CP-19 and CP-21. Resultantly, FCR was significantly (P < 0.05) improved in birds fed with CP-21. Similarly, body weight, feed intake, and FCR were significantly (P < 0.05) improved in enzyme-treated birds compared to the untreated birds. For practical purpose, birds fed CP-19 with protease enzyme resulted in improved FCR compared to birds fed with CP-21. The results revealed that CP and apparent metabolizable energy (AME) were significantly (P < 0.05) higher in birds fed with CP-21 along with supplementation of enzyme while abdominal fat was significantly (P < 0.05) lower in the same group compared to birds fed with CP-17. These results indicated that a diet having 21% CP supplemented with 30,000 IU/kg protease enzyme was superior in improving the performance and digestibility of nutrients in broiler during the starter phase.

Synthetic Kavalactone Analogues with Increased Potency and Selective Anthelmintic Activity against Larvae of Haemonchus contortus In Vitro.

Kava extract, an aqueous rhizome emulsion of the plant Piper methysticum, has been used for centuries by Pacific Islanders as a ceremonial beverage, and has been sold as an anxiolytic agent for some decades. Kavalactones are a major constituent of kava extract. In a previous investigation, we had identified three kavalactones that inhibit larval development of Haemonchus contortus in an in vitro-bioassay. In the present study, we synthesized two kavalactones, desmethoxyyangonin and yangonin, as well as 17 analogues thereof, and evaluated their anthelmintic activities using the same bioassay as employed previously. Structure activity relationship (SAR) studies showed that a 4-substituent on the pendant aryl ring was required for activity. In particular, compounds with 4-trifluoromethoxy, 4-difluoromethoxy, 4-phenoxy, and 4-N-morpholine substitutions had anthelmintic activities (IC50 values in the range of 1.9 to 8.9 µM) that were greater than either of the parent natural products-desmethoxyyangonin (IC50 of 37.1 µM) and yangonin (IC50 of 15.0 µM). The synthesized analogues did not exhibit toxicity on HepG2 human hepatoma cells in vitro at concentrations of up to 40 µM. These findings confirm the previously-identified kavalactone scaffold as a promising chemotype for new anthelmintics and provide a basis for a detailed SAR investigation focused on developing a novel anthelmintic agent.

Epidemiology and antibiogram of common mastitis-causing bacteria in Beetal goats.

BACKGROUND AND AIM: Mastitis has been identified as the most prevalent and economically imperative disease among dairy animals. Thus, understanding its common bacterial pathogens and risk factors is necessary to improve udder health at herd, region, or country level. However, scientific research on caprine mastitis, especially on Beetal breed, has remained to be insufficient in Pakistan. Therefore, this study aimed to evaluate the epidemiology and antibiogram assay of common mastitis-causing bacterial agents, that is, Staphylococcus, Streptococcus, and Escherichia coli, in dairy goats. MATERIALS AND METHODS: In total, 500 Beetal goats, irrespective of age and those that were not treated with any kind of antimicrobial agents during the past 120 h, were screened using California Mastitis Test in Pattoki, Kasur District, whereas epidemiological factors were recorded. The milk samples of mastitic goats were then collected and processed using standard methods. Each sample was primarily cultured on nutrient agar. Using a specific medium, each bacterial colony was separated using several streak methods. Six antibiotic disks belonging to different antibiotic groups were used for antibiogram profiling of bacterial isolates. Chi-square test was used to assess the association of baseline characteristics and mastitis occurrence. Meanwhile, multivariable logistic regression (p<0.001) was utilized to determine the risk factors associated with positive and negative dichotomous outcome of mastitis. RESULTS: The results revealed that the overall prevalence of goat mastitis was 309 (61.8%), in which 260 (52%) and 49 (9.8%) cases were positive for subclinical mastitis (SCM) and clinical mastitis (CM), respectively. Streptococcus and E. coli were found to be the predominant isolates causing SCM and CM, respectively (p<0.001). It was observed that amoxicillin+clavulanic acid was highly sensitive to isolates of Staphylococcus and Streptococcus and ceftiofur sodium to isolates of Streptococcus and E. coli., while enrofloxacin was found to be sensitive to isolates of Streptococcus and E. coli. Risk factors such as herd structure, deworming, vaccination, presence of ticks, use of teat dip and mineral supplements, feeding type, age, parity, housing, blood in the milk, milk leakage, milk taste, and milk yield were found to have the strongest association with mastitis occurrence, while ease of milking has moderate association. CONCLUSION: In the area examined, cases of SCM were found to be higher compared with that of CM, and ceftiofur sodium has been identified as the preferred treatment in both clinical and subclinical forms of caprine mastitis in Beetal goats. Risk factors for mastitis that was identified in this study can form the basis for the creation of an udder health control program specific for dairy goats. We hope our findings could raise awareness of the risk factors and treatment approaches for common mastitis-causing bacterial agents.

Bacterial Spectrum and Antimicrobial Profile of Pediatric Blood Stream Infection at a Tertiary Care Hospital in Pakistan.

Objectives Blood stream infections (BSIs) are the main cause of morbidity and mortality in children worldwide. The present study was carried out to determine the prevalence of BSI with a focus on the identification of the causative agent of BSI, and to further evaluate the antibiotic susceptibility profile of the causing bacterial pathogens.Methods A cross-section study was carried out at the tertiary care hospital in Peshawar, Pakistan from January to December, 2018. Blood samples were collected in BACTECTM bottles and standard microbiological protocols were applied for the isolation and identification of bacterial strains. The antibiotic susceptibility tests were performed using disc diffusion method as per the 2014 guideline of Clinical Laboratory Standard Institute (CLSI).Results Of 567 blood cultures in total, 111(19.6%) were positive for BSI. Male children were 64 % (71/111) and female children were 36% (40/111). For the causative predominant group of microorganisms, Gram-negative bacteria were identified in 79 (71.1%) isolates, and Gram-positive bacteria in 32 (28.9%) isolates. The common bacteria of isolates were S. typhi (n=35, 31.5%), E. coli (n=19, 17.1%), S. aureus (n=18, 16.2%), K. pneumonia (n=12, 10.8%), and Enterococcus species (n=7, 6.3%). The 36.7% (29/79) isolates of Gram-negative bacteria were ESBL producers, and 61.1% (11/18) of S. aureus isolates were methicillin resistant. Overall, 72.9% isolates were multidrug resistant.Conclusions Gram-negative bacteria were the main cause of pediatric BSIs, where the predominant microorganism was S. typhi. Remarkably, majority of the S. typhi isolates were resistant to ciprofloxacin.

Whole genome sequencing of drug resistant Mycobacterium tuberculosis isolates from a high burden tuberculosis region of North West Pakistan.

Tuberculosis (TB), caused by Mycobacterium tuberculosis bacteria, is a leading infectious cause of mortality worldwide, including in Pakistan. Drug resistant M. tuberculosis is an emerging threat for TB control, making it important to detect the underlying genetic mutations, and thereby inform treatment decision making and prevent transmission. Whole genome sequencing has emerged as the new diagnostic to reliably predict drug resistance within a clinically relevant time frame, and its deployment will have the greatest impact on TB control in highly endemic regions. To evaluate the mutations leading to drug resistance and to assess for evidence of the transmission of resistant strains, 81 M. tuberculosis samples from Khyber Pakhtunkhwa province (North West Pakistan) were subjected to whole genome sequencing and standard drug susceptibility testing for eleven anti-TB drugs. We found the majority of M. tuberculosis isolates were the CAS/Delhi strain-type (lineage 3; n = 57; 70.4%) and multi-drug resistant (MDR; n = 62; 76.5%). The most frequent resistance mutations were observed in the katG and rpoB genes, conferring resistance to isoniazid and rifampicin respectively. Mutations were also observed in genes conferring resistance to other first and second-line drugs, including in pncA (pyrazinamide), embB (ethambutol), gyrA (fluoroquinolones), rrs (aminoglycosides), rpsL, rrs and giB (streptomycin) loci. Whilst the majority of mutations have been reported in global datasets, we describe unreported putative resistance markers in katG, ethA (ethionamide), gyrA and gyrB (fluoroquinolones), and pncA. Analysis of the mutations revealed that acquisition of rifampicin resistance often preceded isoniazid in our isolates. We also observed a high proportion (17.6%) of pre-MDR isolates with fluoroquinolone resistance markers, potentially due to unregulated anti-TB drug use. Our isolates were compared to previously sequenced strains from Pakistan in a combined phylogenetic tree analysis. The presence of lineage 2 was only observed in our isolates. Using a cut-off of less than ten genome-wide mutation differences between isolates, a transmission analysis revealed 18 M. tuberculosis isolates clustering within eight networks, thereby providing evidence of drug-resistant TB transmission in the Khyber Pakhtunkhwa province. Overall, we have demonstrated that drug-resistant TB isolates are circulating and transmitted in North West Pakistan. Further, we have shown the usefulness of whole genome sequencing as a diagnostic tool for characterizing M. tuberculosis isolates, which will assist future epidemiological studies and disease control activities in Pakistan.

Phenotypic screening of the 'Kurz-box' of chemicals identifies two compounds (BLK127 and HBK4) with anthelmintic activity in vitro against parasitic larval stages of Haemonchus contortus.

BACKGROUND: Due to anthelmintic resistance problems, there is a need to discover and develop new drugs for the treatment and control of economically important and pathogenic nematodes of livestock animals. With this focus in mind, we screened 236 compounds from a library (called the 'Kurz-box') representing chemically diverse classes such as heterocyclic compounds (e.g. thiazoles, pyrroles, quinolines, pyrimidines, benzo[1,4]diazepines), hydoxamic acid-based metalloenzyme inhibitors, peptidomimetics (bis- and tris-pyrimidoneamides, alkoxyamides) and various intermediates on Haemonchus contortus, one of the most important parasitic nematodes of ruminants. METHODS: In the present study, we tested these compounds, and measured the inhibition of larval motility and development of exsheathed third-stage (xL3) and fourth-stage (L4) larvae of H. contortus using an optimised, whole-organism phenotypic screening assay. RESULTS: Of the 236 compounds, we identified two active compounds (called BLK127 and HBK4) that induced marked phenotypic changes in the worm in vitro. Compound BLK127 induced an 'eviscerated' phenotype in the xL3 stage and also inhibited L4 development. Compound HBK4 exerted a 'curved' phenotype in both xL3s and L4s. CONCLUSIONS: The findings from this study provide a basis for future work on the chemical optimisation of these compounds, on assessing the activity of optimised compounds on adult stages of H. contortus both in vitro and in vivo (in the host animal) and against other parasitic worms of veterinary and medical importance.

Selected α-pyrones from the plants Cryptocarya novoguineensis (Lauraceae) and Piper methysticum (Piperaceae) with activity against Haemonchus contortus in vitro.

Due to the widespread occurrence and spread of anthelmintic resistance, there is a need to develop new drugs against resistant parasitic nematodes of livestock animals. The Nobel Prize-winning discovery and development of the anti-parasitic drugs avermectin and artemisinin has renewed the interest in exploring natural products as anthelmintics. In the present study, we screened 7500 plant extracts for in vitro-activity against the barber's pole worm, Haemonchus contortus, a highly significant pathogen of ruminants. The anthelmintic extracts from two plants, Cryptocarya novoguineensis and Piper methysticum, were fractionated by high-performance liquid chromatography (HPLC). Subsequently, compounds were purified from fractions with significant biological activity. Four α-pyrones, namely goniothalamin (GNT), dihydrokavain (DHK), desmethoxyyangonin (DMY) and yangonin (YGN), were purified from fractions from the two plants, GNT from C. novoguineensis, and DHK, DMY and YGN (= kavalactones) from P. methysticum. The three kavalactones induced a lethal, eviscerated (Evi) phenotype in treated exsheathed third-stage larvae (xL3s), and DMY and YGN had moderate potencies (IC50 values of 31.7 ±â€¯0.23 µM and 23.7 ±â€¯2.05 µM, respectively) at inhibiting the development of xL3s to fourth-stage larvae (L4s). Although GNT had limited potency (IC50 of 200-300 µM) at inhibiting L4 development, it was the only compound that reduced L4 motility (IC50 of 6.25-12.50 µM). The compounds purified from each plant affected H. contortus in an irreversible manner. These findings suggest that structure-activity relationship studies of α-pyrones should be pursued to assess their potential as anthelmintics.

An assessment of worm control practices used by alpaca farmers in Australia.

This study aimed to assess current worm control practices used by Australian alpaca farmers with an online questionnaire survey. The questionnaire contained questions about farm demography and general husbandry practices, farmers' knowledge about gastrointestinal nematodes (GINs) and their importance, the use of worm control strategies and anthelmintics, and grazing management. A link for the questionnaire survey was sent to all (n = 954) registered members of the Australian Alpaca Association in July 2015. The response rate for the questionnaire was 25% (239/954). The majority of respondents were from small (≤ 50 alpacas; 64%, 153/239) followed by medium (50-100 alpacas; 24%, 57/239) and large (>100 alpacas; 12%, 29/239) farms. Findings revealed that the majority of respondents kept Huacaya alpacas to produce high-quality fibre and alpacas were usually kept with other domestic ruminants (e.g. cattle and sheep). Although half of alpaca farmers (114/220) perceived that GINs were an important health problem of alpacas, with Haemonchus spp. being the most common nematode, the majority of them (174/220) used anthelmintics for nematode control. Macrocyclic lactones, a commercial combination of four anthelmintics (abamectin, albendazole, closantel and levamisole) and monepantel were the three most commonly used dewormers by Australian alpaca farmers. Although a significant proportion (166/213) of respondents used a quarantine drench for alpacas, very few respondents were aware of strategic deworming and the issue of anthelmintic resistance. Alpaca farmers mostly used anthelmintics at the dose rate recommended for sheep (47%, 79/167) and cattle (9%, 15/167), though some used 1.5 (31%, 51/167) and 2 (13%, 22/167) times the dose rate recommended for sheep. The majority of small herds used anthelmintics at the dose rate recommended for sheep and cattle while medium and large herds used anthelmintics at 1.5 to 2 times the dose rate recommended for sheep. This study provides invaluable insights into the demography of alpaca farms in Australia, husbandry practices used by alpaca farmers and their knowledge about worms and their control, thereby paving the way for developing guidelines for the control of GINs of alpacas.

Optimization of Novel 1-Methyl-1 H-Pyrazole-5-carboxamides Leads to High Potency Larval Development Inhibitors of the Barber's Pole Worm.

A phenotypic screen of a diverse library of small molecules for inhibition of the development of larvae of the parasitic nematode Haemonchus contortus led to the identification of a 1-methyl-1 H-pyrazole-5-carboxamide derivative with an IC50 of 0.29 µM. Medicinal chemistry optimization targeted modifications on the left-hand side (LHS), middle section, and right-hand side (RHS) of the scaffold in order to elucidate the structure-activity relationship (SAR). Strong SAR allowed for the iterative and directed assembly of a focus set of 64 analogues, from which compound 60 was identified as the most potent compound, inhibiting the development of the fourth larval (L4) stage with an IC50 of 0.01 µM. In contrast, only 18% inhibition of the mammary epithelial cell line MCF10A viability was observed, even at concentrations as high as 50 µM.

Arylpyrrole and fipronil analogues that inhibit the motility and/or development of Haemonchus contortus in vitro.

Due to widespread drug resistance in parasitic nematodes, there is a need to develop new anthelmintics. Given the cost and time involved in developing a new drug, the repurposing of known chemicals can be a promising, alternative approach. In this context, we tested a library (n = 600) of natural product-inspired pesticide analogues against exsheathed third stage-larvae (xL3s) of Haemonchus contortus (barber's pole worm) using a whole-organism, phenotypic screening technique that measures the inhibition of motility and development in treated larvae. In the primary screen, we identified 32 active analogues derived from chemical scaffolds of arylpyrrole or fipronil. The seven most promising compounds, selected based on their anthelmintic activity and/or limited cytotoxicity, are arylpyrroles that reduced the motility of fourth-stage larvae (L4s) with significant potency (IC50 values ranged from 0.04 ±â€¯0.01 µM to 4.25 ±â€¯0.82 µM, and selectivity indices ranged from 10.6 to 412.5). Since the parent structures of the active compounds are uncouplers of oxidative phosphorylation, we tested the effect of selected analogues on oxygen consumption in xL3s using the Seahorse XF24 flux analyser. Larvae treated with the test compounds showed a significant increase in oxygen consumption compared with the untreated control, demonstrating their uncoupling activity. Overall, the results of the present study have identified natural product-derived molecules that are worth considering for chemical optimisation as anthelmintic drug leads.

Screening of the 'Open Scaffolds' collection from Compounds Australia identifies a new chemical entity with anthelmintic activities against different developmental stages of the barber's pole worm and other parasitic nematodes.

The discovery and development of novel anthelmintic classes is essential to sustain the control of socioeconomically important parasitic worms of humans and animals. With the aim of offering novel, lead-like scaffolds for drug discovery, Compounds Australia released the 'Open Scaffolds' collection containing 33,999 compounds, with extensive information available on the physicochemical properties of these chemicals. In the present study, we screened 14,464 prioritised compounds from the 'Open Scaffolds' collection against the exsheathed third-stage larvae (xL3s) of Haemonchus contortus using recently developed whole-organism screening assays. We identified a hit compound, called SN00797439, which was shown to reproducibly reduce xL3 motility by ≥ 70%; this compound induced a characteristic, "coiled" xL3 phenotype (IC50 = 3.46-5.93 µM), inhibited motility of fourth-stage larvae (L4s; IC50 = 0.31-12.5 µM) and caused considerable cuticular damage to L4s in vitro. When tested on other parasitic nematodes in vitro, SN00797439 was shown to inhibit (IC50 = 3-50 µM) adults of Ancylostoma ceylanicum (hookworm) and first-stage larvae of Trichuris muris (whipworm) and eventually kill (>90%) these stages. Furthermore, this compound completely inhibited the motility of female and male adults of Brugia malayi (50-100 µM) as well as microfilariae of both B. malayi and Dirofilaria immitis (heartworm). Overall, these results show that SN00797439 acts against genetically (evolutionarily) distant parasitic nematodes i.e. H. contortus and A. ceylanicum [strongyloids] vs. B. malayi and D. immitis [filarioids] vs. T. muris [enoplid], and, thus, might offer a novel, lead-like scaffold for the development of a relatively broad-spectrum anthelmintic. Our future work will focus on assessing the activity of SN00797439 against other pathogens that cause neglected tropical diseases, optimising analogs with improved biological activities and characterising their targets.