Genomic analyses of a livestock pest, the New World screwworm, find potential targets for genetic control programs.

The New World Screwworm fly, Cochliomyia hominivorax, is a major pest of livestock in South America and Caribbean. However, few genomic resources have been available for this species. A genome of 534 Mb was assembled from long read PacBio DNA sequencing of DNA from a highly inbred strain. Analysis of molecular evolution identified 40 genes that are likely under positive selection. Developmental RNA-seq analysis identified specific genes associated with each stage. We identify and analyze the expression of genes that are likely important for host-seeking behavior (chemosensory), development of larvae in open wounds in warm-blooded animals (heat shock protein, immune response) and for building transgenic strains for genetic control programs including gene drive (sex determination, germline). This study will underpin future experiments aimed at understanding the parasitic lifestyle of the screwworm fly and greatly facilitate future development of strains for efficient systems for genetic control of screwworm.

First report of myiasis in dogs caused by Cochliomyia hominivorax (Coquerel 1858) in Colombia.

This paper reports for the first time in Colombia, two myasis cases caused by Cochliomyia hominivorax in stray dogs. Clinical examination of both patients revealed fever and a large wound at the palpebral level on the right side, and multiple perforating wounds in the right posterior limb where larvae were found. The treatment applied was the manual removal of larvae in the dogs, and subsequent eye enucleation in one dog due to the total loss of the upper right eyelid and the severe mechanical damage that the cornea received. Treatment was reinforced with repellents applied around the wounds and antibiotics. This finding of Cochliomyia hominivorax in stray dogs in Colombia raises concerns about the potential of dogs serving as a reservoir for human infection in the region.

Bovine freeze-dried blood as alternative for feeding of screwworm larvae / Sangre liofilizada bovina como alternativa para la alimentación de larvas del gusano barrenador del ganado

The myiasis caused by Cochliomyia hominivorax (Coquerel) or screwworm is considered one of the most traumatic and relevant to the region due to its economic implications and its importance in public health. The purpose of this study was to compare the reproductive and developmental indexes of screwworm using citrated and freeze-dried bovine blood, spray-dried bovine blood (currently used by Panama-United States Commission for the Eradication and Prevention of Screwworm ñacronym in spanish [Copeg]) and bovine citrated liquid blood. The use of freeze-dried arises through the premise that, due to its process of elaboration constitutes an alternative of better quality for the feeding of the screwworm in function of the integrity of sanguine proteins during its processing. For the comparison of the different diets, the following parameters were used: pH, total pupae weight, pupae size, total pupae volume, emergency percentage, oviposition percentage and hatching percentage. In the case of freeze-dried blood, it showed better mean (standard deviation) indexes of emergence, 83.42 (11.45), and 73.11 (15.97) (p = .01); and hatching, 89.70 (8.88), and 84.08 (8.29), (p = .004) compared to the diet based on spray-dried blood. By virtue of the above, freeze-dried blood proved to be a source of protein with potential to develop insect breeding programs.

La miasis causada por Cochliomyia hominivorax (Coquerel) o gusano barrenador del ganado (GBG) es considerada una de las más traumáticas y relevantes para la región debido a sus implicaciones económicas y su importancia en la salud pública. El propósito de este estudio fue comparar los índices reproductivos y de desarrollo del GBG con sangre bovina liofilizada citratada, sangre bovina secada por pulverización (actualmente utilizada por la Comisión Panamá-Estados Unidos para la Erradicación y Prevención del Gusano Barrenador del Ganado [Copeg]) y sangre líquida bovina citrata. El uso de liofilizado surge bajo la premisa de que, debido a su proceso de elaboración, constituye una alternativa de mejor calidad para la alimentación del GBG en función de la integridad de las proteínas sanguíneas durante su procesamiento. Para la comparación de las diferentes dietas, se utilizaron los siguientes parámetros: pH, peso total de las pupas, tamaño de pupas, volumen total de pupas, porcentaje de emergencia, porcentaje de oviposición y porcentaje de eclosión. En el caso de la sangre liofilizada, mostró mejores promedios (desviación estándar) de índices de emergencia, 83.42 (11.45) y 73.11 (15.97) (p = .01), y eclosión, 89.70 (8.88) y 84.08 (8.29) (p = .004) en comparación con la dieta basada en sangre secada por pulverización. En virtud de lo anterior, la sangre bovina liofilizada demostró ser una fuente de proteína con potencial para desarrollar programas de crianza de insectos.

Epidemiological analysis of the New World screwworm (Cochliomyia hominivorax) in Ecuador.

The New World screwworm (Cochliomyia hominivorax) is an obligate parasite that affects warm-blooded animals. It causes myiasis in livestock and humans, which is a problem for animal production and public health. The health and economic burden of myiasis on livestock production is largely unknown in Ecuador. We investigated the presence of the screwworm and analysed the epidemiology and spatial and temporal trends of myiasis in cattle farms of San Miguel de Los Bancos county. In total, epidemiological questionnaires were conducted in 110 farms, which were subsequently monitored for 12 months. The findings show that the initial and final prevalences in farms were 70% and 61.81%, respectively, and the average monthly prevalence was 15.08%. The initial and final prevalences in animals were 3.87% and 4.60% for bovines and 2.91% and 3.36% for all animals examined. The average percentage of new cases reported per month was 17.68% with a minimum of 10 and a maximum of 28 cases in October and May 2015, respectively. The cumulative incidence estimated that the risk for non-infested farms to become infested could reach 100% in approximately 6 months. The incidence rate is 168 per 1,000 farms at risk-monthly. The annual incidence was 459 per 10,000 for bovines at risk-annually. An analysis of hotspots based on the Getis-Ord Gi* index revealed no temporally stable hot spot, but one temporally stable cold spot, suggesting that most of the study area is generally favourable to infestation, except one cluster of farms.

Wound myiasis: the role of entodermoscopy

Abstract: Although wound or traumatic myiasis is common in tropical countries, only recently cases associated with underlying dermatoses, such as seborrheic dermatitis and psoriasis, have been reported. We describe a patient with seborrheic dermatitis and an ulcerated lesion on the scalp, in which the dermatological examination with the aid of dermoscopy allowed the identification of larvae (maggots) compatible with infestation by Cochliomyia hominivorax. Treatment was performed with oral and topical ivermectin, followed by manual extraction of the larvae.

Wound myiasis: the role of entodermoscopy.

Although wound or traumatic myiasis is common in tropical countries, only recently cases associated with underlying dermatoses, such as seborrheic dermatitis and psoriasis, have been reported. We describe a patient with seborrheic dermatitis and an ulcerated lesion on the scalp, in which the dermatological examination with the aid of dermoscopy allowed the identification of larvae (maggots) compatible with infestation by Cochliomyia hominivorax. Treatment was performed with oral and topical ivermectin, followed by manual extraction of the larvae.

Case Report: Myiasis due to Cochliomyia hominivorax and Dermatobia hominis: Clinical and Pathological Differences between Two Species in Northern Peru.

Infestations caused by fly larvae (Myiasis) have been observed in patients with risk factors and in tropical zones. The aim of our study was to describe the clinical and epidemiological aspects and the risk factors associated with the occurrence of obligatory myiasis, as well as the therapeutic approach to patients. We identified the cases of myiasis diagnosed in two referral hospitals in northern Peru from January 2012 to December 2015 and included patients in whom larval development and a compatible clinical profile were observed. Epidemiological, clinical, analytical, diagnostic, therapeutic, and follow-up data were collected from clinical files. Nine clinical cases were compatible with a diagnosis of myiasis; of these, two were pediatric patients, one was a middle-aged adult, and six were elderly patients. Four of the nine patients were male. The identified species were Dermatobia hominis and Cochliomyia hominivorax. The therapeutic approach included antiparasitic therapy with ivermectin, antibacterial, and in some cases, anti-inflammatory drugs. Dermatobia hominis and C. hominivorax are the predominant species causing myiasis in northern Peru.

Caracterização bioquímica do produto de excreção/ secreção de larvas de Cochliomyia hominivorax (Diptera: Calliphoridae) / Biochemistry characterization of excretion / secretion product of Cochliomyia hominivorax larvae (Diptera: Calliphoridae)

The species Cochliomyia hominivorax, also known as screwworm fly, is an obligate parasite of warm- blooded animals and its geographic range extends thoughout South America, except Chile. This fly causes significant economic losses and has great importance in Brazil. Few studies have focused on the excretion and secretion products of this species, and this research aimed to study the enzymes present in the secretion and excretion (E/S) products of the three larval instars of C. hominivorax. The E/S profile of proteins was obtained by polyacrylamide gel electrophoresis and proteolytic activity was analyzed using gelatin, azocasein and Na-benzoyl-arginine-nitroanilide as substrates.  In E/S products of the three instars, proteins were detected with an apparent molecular weight ranging between 116 and 20 kDa. In the azocasein assay, at different pH ranges, the major proteolytic activity occurred at pH 7.5 for all larval instars. Assays were performed using the same substrates   in which the samples were treated with the inhibitors Benzamidine, Pepstatin A, 4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF), N--tosyl-L-lysine chloromethyl ketone (TLCK), N-- tosyl-L-phenylalanine chloromethyl ketone (TPCK), Ethylenediamine tetraacetic acid (EDTA), and Leupeptin-trans-Epoxysuccinyl-leucylamido(4-guanidino) butane (E-64). Proteinases present in the E/S product of first larvae instar are mostly serine trypsin and chymotrypsin proteases, whereas for second and third instars serine proteases and aspartyl proteases were predominantly observed. Biochemical characterization of E/S products of all larval stages of C. hominivorax helps to improve the understanding of the physiology and the interaction of this parasite with host tissues.

A espécie Cochliomyia hominivorax, conhecida popularmente como mosca da bicheira, é um parasita obrigatório de animais de sangue quente e sua distribuição geográfica estende-se por toda a América do Sul, excetuando-se o Chile. O parasitismo por esta mosca provoca perdas econômicas significativas e tem grande importância no Brasil. São poucos os estudos com foco nos produtos de excreção e secreção desta espécie e este trabalho teve como objetivo estudar as enzimas presentes  no produto de secreção e excreção (E/S) dos três estádios larvais de C. hominivorax. O perfil de proteínas foi obtido por eletroforese em gel de poliacrilamida e a atividade proteolítica foi analisada utilizando-se gelatina, azocaseína e Na-benzoil-arginina-nitroanilida (BAPNA) como substrato. Nos produtos de E/S dos três estádios, as proteínas foram detectadas com um peso molecular aparente que variou entre 116 e 20 kDa. No ensaio de azocaseína, em diferentes faixas de pH, a maior atividade proteolítica ocorreu em pH 7,5 para todos os estádios larvais. Os ensaios foram realizados usando- se estes mesmos substratos e as amostras foram tratadas com os inibidores Benzamidina, Pepstatin A, 4-(2-aminoetil)benzenosulfonil fluoreto hidrocloreto (AEBSF), N--tosil-L-lisina clorometil cetona (TLCK), N--tosil-L-fenilalanina clorometil cetona (TPCK), Ácido etilenodiamino tetra acético (EDTA), Leupeptina e Trans-epoxyysuccinyl L-leucylamido-4-guanidino butano (E-64). As proteinases presentes nos produtos E/S de L1 são em sua maioria serina proteases do tipo tripsina e quimotripsina, enquanto que para os produtos E/S de L2 e L3 foi evidenciada a presença de serinaproteases e aspartil proteases.

Caracterização bioquímica do produto de excreção/ secreção de larvas de Cochliomyia hominivorax (Diptera: Calliphoridae) / Biochemistry characterization of excretion / secretion product of Cochliomyia hominivorax larvae (Diptera: Calliphoridae)

The species Cochliomyia hominivorax, also known as screwworm fly, is an obligate parasite of warm- blooded animals and its geographic range extends thoughout South America, except Chile. This fly causes significant economic losses and has great importance in Brazil. Few studies have focused on the excretion and secretion products of this species, and this research aimed to study the enzymes present in the secretion and excretion (E/S) products of the three larval instars of C. hominivorax. The E/S profile of proteins was obtained by polyacrylamide gel electrophoresis and proteolytic activity was analyzed using gelatin, azocasein and Na-benzoyl-arginine-nitroanilide as substrates.  In E/S products of the three instars, proteins were detected with an apparent molecular weight ranging between 116 and 20 kDa. In the azocasein assay, at different pH ranges, the major proteolytic activity occurred at pH 7.5 for all larval instars. Assays were performed using the same substrates   in which the samples were treated with the inhibitors Benzamidine, Pepstatin A, 4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF), N--tosyl-L-lysine chloromethyl ketone (TLCK), N-- tosyl-L-phenylalanine chloromethyl ketone (TPCK), Ethylenediamine tetraacetic acid (EDTA), and Leupeptin-trans-Epoxysuccinyl-leucylamido(4-guanidino) butane (E-64). Proteinases present in the E/S product of first larvae instar are mostly serine trypsin and chymotrypsin proteases, whereas for second and third instars serine proteases and aspartyl proteases were predominantly observed. Biochemical characterization of E/S products of all larval stages of C. hominivorax helps to improve the understanding of the physiology and the interaction of this parasite with host tissues.(AU)

A espécie Cochliomyia hominivorax, conhecida popularmente como mosca da bicheira, é um parasita obrigatório de animais de sangue quente e sua distribuição geográfica estende-se por toda a América do Sul, excetuando-se o Chile. O parasitismo por esta mosca provoca perdas econômicas significativas e tem grande importância no Brasil. São poucos os estudos com foco nos produtos de excreção e secreção desta espécie e este trabalho teve como objetivo estudar as enzimas presentes  no produto de secreção e excreção (E/S) dos três estádios larvais de C. hominivorax. O perfil de proteínas foi obtido por eletroforese em gel de poliacrilamida e a atividade proteolítica foi analisada utilizando-se gelatina, azocaseína e Na-benzoil-arginina-nitroanilida (BAPNA) como substrato. Nos produtos de E/S dos três estádios, as proteínas foram detectadas com um peso molecular aparente que variou entre 116 e 20 kDa. No ensaio de azocaseína, em diferentes faixas de pH, a maior atividade proteolítica ocorreu em pH 7,5 para todos os estádios larvais. Os ensaios foram realizados usando- se estes mesmos substratos e as amostras foram tratadas com os inibidores Benzamidina, Pepstatin A, 4-(2-aminoetil)benzenosulfonil fluoreto hidrocloreto (AEBSF), N--tosil-L-lisina clorometil cetona (TLCK), N--tosil-L-fenilalanina clorometil cetona (TPCK), Ácido etilenodiamino tetra acético (EDTA), Leupeptina e Trans-epoxyysuccinyl L-leucylamido-4-guanidino butano (E-64). As proteinases presentes nos produtos E/S de L1 são em sua maioria serina proteases do tipo tripsina e quimotripsina, enquanto que para os produtos E/S de L2 e L3 foi evidenciada a presença de serinaproteases e aspartil proteases.(AU)

Oral myiasis caused by Cochliomyia hominivorax treated with ivermectin and nitrofurazone: case report.

Myiasis is the infestation of tissues and organs of animals and humans by certain dipteran fly larvae. This phenomenon is well-documented, especially among animals and people in tropical and subtropical areas. Oral myiasis is a rare condition that can be caused by several species of dipteran fly larvae. When the larvae are removed from the mouth, the tissues seem to recover with no subsequent complications and no need for further treatment. This article presents a case involving a patient with oral myiasis caused by Cochliomyia hominivorax and treated with ivermectin and nitrofurazone.

Histerocele total complicado con miasis a propósito de un caso / Histerocele myiasis complicated by a total of one case

Miasis es la infestación de órganos o tejidos por larvas de moscas. La infestación con larvas de mosca produce diversas manifestaciones según el sitio afectado y puede incluso, causar la muerte. No es una enfermedad común en humanos pero se observa con alguna regularidad en países neotropicales. Afecta con mayor frecuencia las áreas expuestas de la piel y se presenta raramente en ojos, nariz, senos paranasales, tracto urogenital o recto; en estos casos la infestación se asocia con traumas previos o secresiones purulentas que atraen a las moscas adultas. Se presenta el caso de una paciente femenina de 65 años de edad, con prolapso total del útero (histerocele grado IV) complicado con miasis específica por Cochliomya hominivorax.

Use of polyester pad as a new physical substrate for rearing Cochliomyia hominivorax coquerel (Diptera: Calliphoridae) larvae.

Polyester pad was utilized as solid support for rearing Cochliomyia hominivorax Coquerel larvae in liquid diet and compared with the ground meat diet. There were no significant differences in the mean third instar larvae and pupal weights, number of pupae, emergence and rearing efficiency. The tests were conducted through four consecutive generations, presenting no detectable effect in any of the life history parameters. We propose that the polyester pad can be used as solid support for rearing C. hominivorax larvae in liquid diets and can be reutilized reducing the costs of mass rearing.

Genetic approaches for studying myiasis-causing flies: molecular markers and mitochondrial genomics.

"Myiasis-causing flies" is a generic term that includes species from numerous dipteran families, mainly Calliphoridae and Oestridae, of which blowflies, screwworm flies and botflies are among the most important. This group of flies is characterized by the ability of their larvae to develop in animal flesh. When the host is a live vertebrate, such parasitism by dipterous larvae is known as primary myiasis. Myiasis-causing flies can be classified as saprophagous (free-living species), facultative or obligate parasites. Many of these flies are of great medical and veterinary importance in Brazil because of their role as key livestock insect-pests and vectors of pathogens, in addition to being considered important legal evidence in forensic entomology. The characterization of myiasis-causing flies using molecular markers to study mtDNA (by RFLP) and nuclear DNA (by RAPD and microsatellite) has been used to identify the evolutionary mechanisms responsible for specific patterns of genetic variability. These approaches have been successfully used to analyze the population structures of the New World screwworm fly Cochliomyia hominivorax and the botfly Dermatobia hominis. In this review, various aspects of the organization, evolution and potential applications of the mitochondrial genome of myiasis-causing flies in Brazil, and the analysis of nuclear markers in genetic studies of populations, are discussed.

Evaluation of the prophylactic effect and curative efficacy of fipronil 1% pour on (Topline) on post-castration scrotal myiasis caused by Cochliomyia hominivorax in cattle.

A field trial was carried out during a summer-fall period on a commercial beef cattle farm in Minas Gerais State, located in the Southeast of Brazil. In order to evaluate the prophylactic effect and the curative efficacy of fipronil in a 1% solution, 200 Zebu crossbred bulls, with ages varying from 20 to 30 months and weights from 233 to 362 kg, were selected. The bulls were assigned by ranked pair to an untreated control group (A) or to a treated group (B), resulting in 100 animals per group. All experimental animals were surgically castrated on day 0, following routine procedures. After castration all animals in the group B were treated with 10 mg/kg bw of a 1% fipronil solution, topically on the dorsal mid-line. The wounds were individually inspected on days: 2, 4, 7, 10, 14, 17, 21, 28 and 35. After castration the animals were naturally exposed to Cochliomyia hominivorax and remained in the same pasture throughout the trial. Among the animals in the control group, 83 were observed to harbor C. hominivorax eggs, with a total of 97 ovipositions, and among those 73 animals had active myiasis. In group B (fipronil 1%), 66 animals showed C. hominivorax eggs, with 92 ovipositions and five animals with active myiasis. Most ovipositions and active myiasis were detected until seven days post-castration for both groups. Wound parasite infestation evidenced bleeding, serous purulent exudation and presence of active C. hominivorax larvae. Treatment with fipronil 1% had a prophylactic effect on scrotal wounds against the development of C. hominivorax larvae in more than 95% of the treated animals for up to 17 days after castration. The treatment showed partial protection of 66% and 50% on days 21 and 28 post-treatment (pt), respectively. Three animals from the control group and one from the treated group showed active screwworms on day 21 pt, and one animal from the treated group and two from the control group also presented C. hominivorax larvae on scrotal wounds on day 28 pt. By the end of the observation period (day 35 pt), the castration wound had healed in all animals. All experimental animals presenting scrotal wounds infested with C. hominivorax larvae were treated with a 1% pour-on formulation of fipronil, on the same day that infestation was observed. Active C. hominivorax larvae were not seen during the monitoring period immediately after treatment. The curative efficacy of fipronil 1% against C. hominivorax larvae infestation in castration wounds was 100%.

Orbital myiasis in a patient with a chronically exposed hydroxyapatite implant.

We describe the clinical presentation of the first case, to our knowledge, of myiasis in an orbit with an exposed hydroxyapatite implant. We examined a 10-year-old boy with an 8-year exposure of an orbital implant. He had an infestation with maggots. The implant and the worms were surgically removed, allowing the socket to heal by secondary intention. The patient was able to wear a prosthesis 2 weeks later. The parasites were identified as belonging to Cochliomyia hominivorax, a fly responsible for the majority of cases of myiasis in the Western hemisphere.

Responses of fertile and sterile screwworm (Diptera: Calliphoridae) flies to bovine blood inoculated with bacteria originating from screwworm-infested animal wounds.

ABSTRACT A simple bioassay system was developed to study locomotory and ovipositional responses of screwworm, Cochliomyia hominivorax (Coquerel), flies to bovine blood inoculated with eight species of coliform bacteria that were isolated from screwworm-infested animal wounds. When exposed to odors from bacteria-inoculated blood which was incubated for 72 h at 37 degrees C, approximately 50% of 7- and 10-d-old gravid females landed on the blood by the end of 15 min test exposure. Only 17% of 7-d-old reproductively sterile females (from irradiated pupae) with previtellogenic ovaries and 2% of 4-d-old vitellogenic females responded to the same treatment. Females generally reacted in greatest numbers to bacteria-inoculated blood incubated for 72 h, followed by 48 h, then 24 and 96 h. Males of all ages tested were unresponsive. Although oviposition occurred in tests with gravid females lasting for 1 h, with both inoculated blood and an uninoculated control, the inoculated sample was significantly better than the control at 48, 72, and 96 h incubation duration. Our results are consistent with the conclusion that the inoculated blood, when incubated for 48-72 h, gives off volatile chemicals which attract gravid females and contains an oviposition stimulant that acts following contact and feeding. The volatiles, once isolated and identified, may be useful for sampling gravid females in the field as well as improving the oviposition system in the mass-production facility of the screwworm eradication program.

Efficacy of dicyclanil in the prevention of screwworm infestation (Cochliomyia hominivorax) in cattle castration wounds.

A field trial was carried out during summer 1996 in a commercial beef cattle farm located in the central area of Argentina (30 degrees 16'S, 60 degrees 30'W) to evaluate the efficacy of a new insect growth regulator material (dicyclanil) in the prevention of myiosis caused by screwworm (Cochliomyia hominivorax) larvae. Forty bull calves (Bos indicus x Bos taurus), 5 to 6 months old were allocated by ranked paris to a control (G.1) and a treated group (G.2) of the same number of animals. On day 0, all experimental animals were surgically castrated and calves of group 2 were treated topically with 20 ml as a single dose of a ready-to-use formulation containing 5% w/v dicyclanil. Inspections for screwworm larva infestation were carried out on days 4, 8, 12, 16, 19, 23 and 25 after castration. The incidence of egg masses on the scrotal wounds evidenced a similar challenge for all groups (P > 0.05 chi(2)). However, 16 animals of G.1 (80%) and only one of G.2 (5%) developed active myiosis, respectively. This difference was significant (P=0.003 X(2)).

Myiasis cutáneo-ulcerosas en pequeños animales / Cutaneous myiasis in small animals

En nuestro país los estudios acerca de las myiasis cutáneo-ulcerosas son escasos e incompletos, y de las que afectan a pequeños animales sólo se tienen datos de infecciones inducidas en la provincia de Buenos Aires. En este estudio se detectaron 149 perros y 9 gatos afectados por larvas de Cochliomyia hominivorax, 11 perros y 10 gatos por larvas de Phaenicia sericata. Las lesiones variaron desde superficiales leves hasta cavernosas muy graves, las mismas se hallaron distribuidas en la cabeza (boca, ojos, nariz y orejas), tronco, miembros, genitales externos, ano, periné y base de la cola. Las larvas I, II y III fueron extraídas en su totalidad entre los días 1 y 7 postinfección, en un número que varió entre 1 y 250. Los adultos de C. hominivorax han sido asociados a pasturas y floresta, y se ejerce sobre ellos control y vigilancia en las áreas subtropicales para evitar pérdidas en la producción ganadera. Los hallazgos en pequeños animales de zonas templadas y vinculados a áreas urbanas, alertaría sobre los riesgos de dispersión de una parasitosis de consecuencias aún no evaluadas en nuestro país. Las myiasis por P. sericata están estrechamente relacionadas con tejidos necrosados o cadáveres, por lo tanto, los datos aquí aportados representan los primeros registros de la especie como agente causal de myiasis primaria en pequeños animales