Identification of Cross-Resistance and Multiple Resistance in Eimeria tenella Field Isolates to Commonly Used Anticoccidials in Pakistan

A battery trial was conducted to evaluate the drug sensitivity in Eimeria tenella field isolates against the commonly used anticoccidials salinomycin (60 ppm), maduramicin (5 ppm), and clopidol (125 ppm) in broiler chicks. These anticoccidials were mixed in feed at d 12 of age, and inoculation was given on d 14 of age. Drug sensitivity was determined by using the global index, which is composed of percentage weight gain, FCR, lesion score, oocyst index, and mortality percentage. In the present study, all the E. tenella isolates showed partial resistance against salinomycin, whereas varying degrees of sensitivity were observed against maduramicin and clopidol.     

Efficacy of decoquinate against drug sensitive laboratory strains of Eimeria tenella and field isolates of Eimeria spp. in broiler chickens in China

The efficacy of decoquinate against Eimeria infections in broiler chickens was evaluated using two drug sensitive laboratory strains of Eimeria tenella and 20 field isolates of Eimeria spp. collected from farms in China where various anticoccidials (including maduramicin) had been used. Decoquinate (20-40 ppm in feed) and maduramicin (5 ppm) were efficacious against E. tenella laboratory strains, but decoquinate more so than maduramicin. Body weight gains of E. tenella infected chickens were significantly improved, and caecal lesions were prevented, by feeding either decoquinate or maduramicin. Decoquinate also prevented oocyst production, but maduramicin did not. Most (18/20) Eimeria field isolates were resistant to maduramicin, judged by oocyst production; decoquinate at > or =20 ppm completely controlled all 20 field isolates. Decoquinate has potential value as a broiler anticoccidial in China and other countries where it has not been previously used.     

Efficacy of maduramicin against ionophore-tolerant field isolates of coccidia in broilers

Maduramicin ammonium was given at 2.5-8 ppm in the feed to broilers experimentally infected with coccidia recently isolated from broiler farms where ionophores had been used for several years. Infection pressure varied from mild to severe in five trials: mortality in unmedicated controls ranged from 0 to 59%, intestinal lesion scores were high, and weight gain was depressed by the infections. The cultures of Eimeria were partly resistant to ionophores: birds medicated with monensin at 100-121 ppm had only modest reductions in lesion scores and incomplete protection against weight loss or mortality. Control of infections by maduramicin was significant at 4 ppm but best at 5-7 ppm. Maduramicin was more effective than monensin or narasin, but about the same as salinomycin, in reducing lesions and mortality and in protecting performance. Maduramicin was well tolerated within the dose range of 5-7 ppm.     

The efficacy of anticoccidial products against Eimeria spp. in northern bobwhites

To determine whether chemotherapeutic compounds available for use in domestic poultry are effective at controlling coccidiosis in northern bobwhites (Colinus virginianus), we tested 13 chemotherapeutic anticoccidials including amprolium (250 parts per million [ppm]), clopidol (125 ppm), diclazuril (1 ppm and 2 ppm), decoquinate (30 ppm), lasalocid (120 ppm), monensin (90 ppm), narasin/nicarbazin (36/36 ppm), robenidine (33 ppm), roxarsone (50 ppm), sulfadimethoxine/ ormetoprin (125/75 ppm), salinomycin (60 ppm), semduramicin (25 ppm), and zoalene (125 ppm and 150 ppm). Three tests were conducted using two replicates of 10 birds each: Infected, unmedicated controls and medicated birds were challenged with 1 x 10(6) oocysts of a field isolate consisting primarily of Eimeria lettyae. Subsequently, we tested clopidol, lasalocid, salinomycin, diclazuril (1 ppm), and monensin against mixed-species field isolates containing E. lettyae, E. dispersa, E. colini, or all. Weight gain, gross intestinal lesions, severity of diarrhea, and feed conversion ratio (FCR) 6 days postinfection were recorded. Lesion score, as previously reported, was unreliable as a measure of severity of infection in comparison with weight gain, fecal scores, and FCR. Excellent to good efficacy was found in clopidol, decoquinate, diclazuril (1 ppm and 2 ppm), and in lasalocid, narasin and nicarbazin, robenidine, sulfadimethoxine/ormetoprin, and zoalene (150 ppm). Marginal protection was found using monensin, salinomycin, semduramicin, or a roxarsone/semduramicin combination. Amprolium, roxarsone, and zoalene (125 ppm) were ineffective at controlling coccidia. Two of the six isolates tested against diclazuril 1 ppm and clopidol demonstrated a high degree of resistance, but none of the six isolates was resistant to lasalocid. Four of the eight isolates showed mild to moderate, and moderate to high, resistance against monensin and salinomycin, respectively. These findings indicate that several available compounds are effective at controlling coccidiosis in bobwhites.     

Isolation and selection of ionophore-tolerant Eimeria precocious lines: E. tenella, E. maxima and E. acervulina

Eimeria parasites were isolated from Nanhai Guangdong province (southern China) and studied in chickens in wire cages to evaluate their drug resistance against commonly used ionophores: monensin (100 mg/kg of feed), lasolacid (90 mg/kg), salinomycin (60 mg/kg), maduramicin (5 mg/kg) and semduramicin (25 mg/kg). Chinese Yellow Broiler Chickens were infected with 40,000 crude sporulated Eimeria oocysts at 15 days of age and prophylactic medication commenced a day prior to infection. Drug resistance was assessed for each ionophore drug by calculating the anticoccidial index (ACI) and percentage optimum anticoccidial activity (POAA) based on relative weight gain, rate of oocyst production and lesion values. Results revealed that Nanhai Eimeria oocysts comprising of E. tenella, E. maxima and E. acervulina, were resistant to monensin, sensitive to both salinomycin and lasolacid and partially sensitive to maduramicin and semduramicin. By selection for early development of oocysts during passage through chickens, the prepatent time of E. tenella, E. maxima and E. acervulina were reduced by 49, 36 and 22 h, respectively. The precocious lines are less pathogenic than the parent strains from which they were selected and conferred a satisfactory protection for chickens against coccidiosis. These ionophore-tolerant precocious lines could have wider applications in the development of anticoccidial vaccines for sustainable control of coccidiosis.     

Efficacy of maduramicin ammonium against coccidiosis in turkeys under laboratory and floor-pen conditions

Experimental infections with field isolates of Eimeria meleagrimitis, E. adenoeides, E. gallopavonis, and E. dispersa in turkey poults were used to test the efficacy of maduramicin ammonium at 2.5-10 ppm in laboratory experiments. Infection with single or mixed species of coccidia reduced the weight gain of unmedicated infected controls and caused 18.1-65% mortality in two experiments. Maduramicin ammonium given at 5-7 ppm prevented mortality, significantly reduced droppings scores and oocyst passage, and improved weight gain to near that of the unmedicated uninfected controls. Maduramicin ammonium was tested at 4-7 ppm in a floor-pen trial lasting 10 weeks. Mortality from coccidiosis averaged 11.9% in unmedicated controls, compared with 0.6% with 4 ppm of maduramicin or no mortality with 5-7 ppm. Average weight gain and feed conversion at 10 weeks were significantly improved over unmedicated infected controls when maduramicin ammonium was given at 5-7 ppm. These results suggest that maduramicin ammonium is highly efficacious against field isolates of Eimeria in turkeys, especially within the range of 5-7 ppm in the feed.     

In vitro efficacy and pharmacodynamic profiles of four polyether ionophores against methicillin-resistant Staphylococcus spp.

The objective of this study was to determine the minimum inhibitory concentrations (MICs) and pharmacodynamic profiles of four ionophores (lasalocid, monensin, narasin and salinomycin) against staphylococcal isolates from clinical cases of human and veterinary staphylococcal infections, and to determine the effect of methicillin resistance on the antimicrobial activity of ionophores. Broth microdilution MIC testing was used to determine antimicrobial activity against 156 staphylococcal isolates of human and veterinary origin. Pharmacodynamic profiles were examined using time-kill kinetics profiles against an ATCC type strain of Staphylococcus aureus and a clinical isolate of methicillin-resistant Staphylococcus pseudintermedius. All tests were performed in accordance with CLSI guidelines. All four ionophores demonstrated antimicrobial activity against methicillin-resistant staphylococci at concentrations similar to those observed for methicillin-susceptible isolates of the same species. Testing of human and veterinary MRSA isolates also showed that MIC values were not influenced by the host origin of the isolates. Pharmacodynamic profiles were similar for both isolates tested across all four ionophores, with similar reductions in viable cell counts being observed over an 18- to 24-hr period. Lasalocid, monensin, narasin and salinomycin all demonstrated antimicrobial activity against staphylococcal isolates of human and veterinary origins, with activity being unaffected by methicillin resistance status, although some Staphylococcus species-specific effects were observed that require further investigation.     

Anticoccidial evaluation of halofuginone, lasalocid, maduramicin, monensin and salinomycin

The activities of five anticoccidials were compared against Eimeria species in/of chickens, in controlled in vivo and in vitro laboratory studies. Two more recent and potent market entries (maduramicin and halofuginone) were compared with three older polyether antibiotic anticoccidials (monensin, lasalocid and salinomycin). Halofuginone, lasalocid, maduramicin, monensin and salinomycin were evaluated at 3, 125, 5, 120 and 66 ppm, respectively, of active drug in the diets. At these levels, all five drugs demonstrated significant activity against Eimeria tenella, E. maxima, E. necatrix, E. brunetti and E. acervulina (in vivo). Monensin was least effective against E. tenella, and one of the lesser efficacious drugs against E. necatrix, maduramicin, was least effective against E. maxima. In studies of single Eimeria species infections, comparable weight gains were noted for the drugs. In the mixed Eimeria species infections, however, birds treated with maduramicin had significantly higher weight gains than did birds medicated with monensin. Unlike in vivo potencies, titration in vitro indicated that monensin was most potent (active at 10(-6) mcg ml-1), and maduramicin and lasalocid least potent (inactive at less than or equal to 10(-3) mcg ml-1).     

A survey of sensitivity to anticoccidial drugs in 60 isolates of coccidia from broiler chickens in Brazil and Argentina

Coccidia were isolated from 90 broiler farms in 15 poultry-producing areas in Brazil and Argentina. Sixty isolates were tested for sensitivity to 7 anticoccidial drugs. The common species were: a) Eimeria tenella, 47 isolates; b) E. maxima, 49 isolates; c) E. acervulina, 44 isolates; d) E. mitis, 26 isolates; and e) E. brunetti, 12 isolates. Isolates were considered sensitive to drugs if intestinal lesion scores of medicated broilers were reduced by at least 50% compared with unmedicated infected broilers or if weight gain was at least 75% of that of uninfected birds in a 6-day laboratory test. According to lesion scores, there was evidence of resistance or seriously reduced sensitivity to monensin in 20 isolates, narasin in 29, salinomycin in 11, maduramicin in 1, clopidol in 36, amprolium in 40, and nicarbazin in 1. According to broiler weight gain, there was resistance to monensin in 36 isolates, narasin in 32, salinomycin in 28, maduramicin in 2, clopidol in 28, amprolium in 50, and nicarbazin in 4. These results suggested incomplete cross resistance of coccidia to polyether ionophorous drugs. The degree of resistance might be explained by previous patterns of use of these drugs.     

堆型艾美耳球虫早熟株对常用抗球虫药的敏感性测定

The study was aimed to explore the sensitivity of Eimeria acervulina （E.acervulina） precocious strain to some common anticoccidials (diclazuril, toltrazuril, sulfamonomethoxine, dinitolmide, decoquinate, nicarbazin, maduramicin, salinomycin and oxytetracycline). The sensitivity of E.acervulina precocious strain to 9 kinds anticoccidials was assessed by using 4 indices: anticoccidial activity percentage (POAA), reduction of lesion scores (RLS), relative oocyst production (ROP) and anticoccidial index (ACI). The results of these sensitivity trials indicated that the four indices POAA, RLS, ROP and ACI of groups toltrazuril, sulfamonomethoxine, dinitolmide, decoquinate, nicarbazin, salinomycin and oxytetracycline were all negative; the ROP index of diclazuril group was positive, the other three indices were negative. The ROP and ACI indices of maduramicin group were positive, the other two indices were negative. The results showed that the Eimeria acervulina precocious strain was sensitive to diclazuril, toltrazuril, sulfamonomethoxine, dinitolmide, decoquinate, nicarbazin, salinomycin and oxytetracycline, it was partly sensitive to maduramicin.     

Cardiomyopathy in cattle induced by residues of the coccidiostat maduramicin in poultry litter given as a feedstuff

An experimental feeding trial was conducted in order to substantiate an hypothesis on the aetiology of a previously unrecorded cardiomyopathic syndrome in beef cattle in Israel. It was believed that residues of the ionophore maduramicin (Cygro; American Cyanamid) in poultry litter fed to cattle, after the maduramicin had been incorporated into broiler feed as a coccidiostat, were the cause of the cardiomyopathy. Three groups of 5 heifers were fed for several weeks (1) poultry litter from a field case, containing 4.8 ppm maduramicin; (2) poultry litter to which maduramicin was added to give a concentration of 12 ppm; (3) poultry litter with no maduramicin (control). Clinical, biochemical, necropsy and histopathological findings showed that maduramicin residues are cardiotoxic, even at the lower level of maduramicin fed, which is commonly encountered in poultry litter in Israel.     

离子载体药物对家鸭球虫病治疗效果比较

鸭球虫病是由多种鸭艾美耳球虫寄生于鸭肠上皮细胞引起的一种原虫病,给养禽业带来巨大的经济损失。为了解离子型抗球虫药物对鸭球虫病的防治效果,在模拟家鸭笼养的环境条件下,选用莫能霉素、盐霉素、马杜拉霉素、拉沙里菌素、那拉霉素等5种离子载体药物进行试验。试验结果表明,5种离子载体类药物对鸭球虫病均有很好的防治效果,并且马拉霉素等对家鸭的增重也有一定效果。     

上海地区鸡球虫对6种抗球虫药的抗药程度研究

为了解上海地区的鸡球虫抗药性状况,作者采集了上海市9个区县10个鸡场的球虫,进行了对地克珠利（Diclazuril)、氯苯胍（Robenidine)、氯羟吡啶（Clopidol)、氨丙啉（Amprolium)、马杜拉霉素（Maduramicin)、莫能霉素（Monensin)的抗药性研究。试验设6个药物组和感染不用药、不感染不用药2个对照组,以抗球虫指数（ACI）、病变记分减少率（RLS）、相对卵囊产量（ROP）、最适抗球虫活性百分率（POAA）为试验指标,进行综合评定,其抗药程度分为无抗药性、轻度抗药性、中度抗药性、完全抗药性4个等级。结果显示来自10个鸡场的球虫中,对地克珠利有90％未产生抗药性,对氯苯胍有90％为无抗药性和轻度抗药,对驻杜拉霉素有50％为无抗药性和轻度抗药,对氯羟吡啶有80％达到完全抗药,对氨丙啉有90％达到中度和完全抗药,对莫能霉素则全部达到中度以上的抗药程度,作者认为目前在上海地区以放心地理合理使用地克珠利和氯苯胍,对马杜拉霉素要慎使用,对氯羟吡啶、氨丙啉、莫能霉素须减少或暂停使用。     

Repurposing Ionophores as novel antimicrobial agents for the treatment of bovine mastitis caused by Gram‐positive pathogens

Increasing reports of multidrug‐resistant bacterial infections in animals has created a need for novel antimicrobial agents that do not promote cross‐resistance to critically important antimicrobial classes used in human medicine. In response to the recent emergence of antimicrobial resistance in several bovine mastitis pathogens, in vitro antimicrobial susceptibility was determined for four polyether ionophores (lasalocid, monensin, narasin and salinomycin) against Staphylococcus spp. and Streptococcus spp. isolated from clinical cases. In addition, erythrocyte haemolysis and WST ‐1 cell proliferation assays were used to assess in vitro mammalian cell cytotoxicity and biofilm susceptibility testing was performed using the minimum biofilm eradication concentration (MBEC ?) biofilm assay. Lasalocid, monensin, narasin and salinomycin exhibited bacteriostatic antimicrobial activity against all pathogens tested, including methicillin‐resistant staphylococci, with MIC ₉₀ values <16 μg/ml. Narasin and monensin displayed the least toxicity against mammalian cell lines and all compounds significantly reduced viable cell numbers in a Staphylococcus aureus biofilm. Based on in vitro characterization, all four ionophores offer potentially novel treatments against bovine mastitis but in vivo studies will be essential to determine whether acceptable safety and efficacy is present following intramammary administration.     

Efficacy of some anticoccidial drugs for treating coccidial enteritis of the common carp caused by Goussia carpelli (Apicomplexa: Eimeriidae)

In this study, nine anticoccidial drugs commonly used in poultry were tested for efficacy for the prevention and treatment of Goussia carpelli (Apicomplexa) infection in common carp (Cyprinus carpio L.). To establish experimental infection with G. carpelli, paratenic host oligochaetes of the genera Tubifex and Limnodrilus were infected with oocysts, and laboratory-cultured parasite-free common carp fingerlings were infected by feeding to them oligochaetes containing sporozoites. The anticoccidial drugs (amprolium, narasin, maduramicin, salinomycin Na, lasalocid Na, diclazuril, robenidine HCl, monensin Na and toltrazuril), mixed in the food of the fish in a dose of 200 mg/kg, were fed for 12 days. Common carp fingerlings fed diclazuril, lasalocid, robenidine HCl or maduramicin and killed on day 14 after exposure were free from infection, while other groups treated with amprolium, toltrazuril, monensin Na, narasin or salinomycin Na harboured oocysts in the mucus and epithelium of the gut.     

巨型艾美耳球虫和柔嫩艾美耳球虫早熟株对8种抗球虫药的敏感试验

为检测巨型艾美耳球虫和柔嫩艾美耳球虫早熟株对常用抗球虫药的敏感性,选用地克珠利、氯苯胍、二硝托胺、癸氧喹酯、尼卡巴嗪、马杜米星、盐霉素和拉沙洛西等8种抗球虫药进行鸡体试验,通过抗球虫活性百分比（percent of optimum anticoccidial aetivity,POAA）、病变记分减少率（reduction of lesion scores,RLS）、相对卵囊产量（relative oocyst production,ROP）和抗球虫指数（anticoccidial index,ACI）4项指标进行综合评价。结果表明,巨型艾美耳球虫早熟株对8种药物敏感;柔嫩艾美耳球虫早熟株对盐霉素轻度敏感,对地克珠利、氯苯胍、二硝托胺、癸氧喹酯、尼卡巴嗪、马杜米星和拉沙洛西敏感。本研究结果可为对球虫早熟株疫苗的开发与应用提供实验依据。     

Drug-sensitivity of 99 isolates of coccidia from broiler farms

Coccidia were isolated from 99 broiler farms in 12 broiler-producing states and tested for sensitivity to contemporary anticoccidial drugs. The isolates usually comprised two or more species, including 92 Eimeria acervulina, 47 E. maxima, 28 E. tenella, and 10 E. brunetti. Based on intestinal-lesion-score reduction of 30% or more, 38% of isolates were resistant to monensin (110 ppm), 29% to salinomycin (60 ppm), 20% to nicarbazin (125 ppm), and 46% to amprolium + ethopabate (125 + 4 ppm). Isolates with 50% or greater reduction of lesion scores were considered sensitive. Thirty-three percent were sensitive to monensin, 53% to salinomycin, 67% to nicarbazin, and 39% to amprolium + ethopabate. Other isolates had intermediate response to drugs and were considered to have "reduced sensitivity." There was some evidence for incomplete cross resistance to the polyether, ionophorous products.     

2株柔嫩艾美耳球虫对5种抗球虫药的抗药性

选择130只21日龄雏鸡，研究了柔嫩艾美耳球虫（E.tenella)南京株和凤阳株对地克珠利（Diclazuril)、马杜霉毒（Maduramicin)、氯羟吡啶（Clopidol)、氯苯胍（Robenidine)和氨丙啉（Amprolium)的抗药性。试验鸡随机分成13组，其中每株球虫均设5个感染用药组，1个感染不用药组，另设1个不感染不用药组，每组10只鸡。以POAA，RLS，ROP，ACI作为指标。结果表明，2株柔嫩艾美耳球虫均对氨丙啉轻度抗药，对地克珠利敏感或轻度抗药，对氯羟吡啶中度抗药。对马杜霉和氯苯胍中度抗药或安全抗药。提示：目前在凤阳地区可合理地使用地克珠利和氨丙啉，对氯羟哟啶应慎用，对马杜霉素和氯苯胍须减少或暂停使用。     

Simultaneous treatment of chickens with salinomycin and tiamulin in feed.

Laboratory and field experiments involving more than 100,000 birds were performed to assess the effect of simultaneous in-feed medication of chickens with salinomycin and tiamulin at various concentrations. In an artificial infection study with Mycoplasma gallisepticum, low levels of tiamulin (10-40 ppm) did not induce signs of ionophore intoxication with salinomycin at 60 ppm in the feed, whereas levels of 50 ppm caused early signs with a mild growth depression. A level of 20 ppm gave a maximum average improvement in growth rate of 12.5%. There was a dose-related response in mycoplasma lesion inhibition, with 50 ppm reducing lesions by 75%. In the two field trials, chickens suffering from mycoplasmosis complicated with infectious bronchitis and receiving 60 ppm salinomycin in the feed showed marked improvements in mortality rates, lesion scores, and feed-conversion efficiency when tiamulin was added at 20 ppm and 30 ppm in the feed. There were no signs of incompatibility between the two antibiotics at these levels.     

5株柔嫩艾美耳球虫对4种抗球虫药的抗药性

选择260只19日龄雏鸡，随机分成26组，每组10只，研究柔嫩艾美耳球虫(E．tenella)南京株、凤阳株、扬州株、宣城株和蒙城株对地克珠利(Diclazuril)、马杜霉素(Maduramicin)、氯羟吡啶(C1opido1)、氯苯胍(Robenidine)的抗药性。每株球虫均设4个感染用药组、1个感染不用药组，另外设1个不感染不用药组。以POAA、RLS、ROP、ACI作为指标，判定5株球虫对4种药物的抗药性。结果表明凤阳株、扬州株和蒙城株分别仅对马杜霉素、氯苯胍、地克珠利敏感，南京株和宣城株对该4种药物均已产生不同程度的抗药性。提示目前在凤阳、扬州和蒙城地区可分别合理地使用马杜霉素、氯苯胍和地克珠利，对其它药物须减少或暂停使用。