Dose determination of a novel formulation of metaflumizone plus amitraz for control of cat fleas (Ctenocephalides felis felis) and brown dog ticks (Rhipicephalus sanguineus) on dogs.

A novel spot-on formulation containing metaflumizone and amitraz (ProMeris/ProMeris Duo for Dogs, Fort Dodge Animal Health, Overland Park, KS) was evaluated in a laboratory study to determine the appropriate dose for efficacy against fleas and ticks on dogs for 1 month. Thirty-six Beagles were randomly allocated to six equal groups and individually housed. One group remained nontreated. Another was treated with a placebo formulation (solvents with no active ingredients). Three groups of dogs were treated topically with the metaflumizone plus amitraz formulation (150mg of each of metaflumizone and amitraz/ml), at volumes providing doses of 10, 20 and 40mgeachactive/kg. The final group was treated with a commercial spot-on providing 6.7mgfipronil/kg. All treatments were applied to the skin at a single spot between the scapulae on Day 0. Dogs were infested with 50 adult brown dog ticks (Rhipicephalus sanguineus) on each of Days -2, 5, 12, 19, 26, 33 and 40, and with 100 cat fleas (Ctenocephalides felis felis) on Days -1, 6, 13, 20, 27, 34 and 41. Dogs were examined and parasites "finger counted" on Day 1 to estimate knock down efficacy, and all animals were comb counted to determine the numbers of viable fleas and ticks on Days 7, 14, 21, 28, 35 and 42. There were no significant differences in parasite counts between the nontreated control and the placebo-treated control groups for either fleas or ticks (P>0.05) except for very slight reductions on Day 7 for fleas and Day 14 for ticks, demonstrating that the formulation excipients had no activity. The qualitative finger counts on Day 1 indicated that all of the insecticidal treatments resulted in a noticeable reduction in flea and tick numbers within 1 day of treatment. All of the metaflumizone and amitraz treatments and fipronil resulted in significantly lower flea and tick numbers relative to nontreated controls on all posttreatment count days (P<0.05). For the metaflumizone plus amitraz treatments, mean flea and tick counts for the 10mg/kg dose were significantly higher than those for the 20mg/kg dose (P<0.05) from Day 21 on. There was no significant advantage provided by the 40mg/kg dose over the 20mg dose throughout the entire study (P>0.05). The two higher metaflumizone plus amitraz doses provided >95% control of fleas and >90% control of ticks for at least 35 days after treatment, and this level of control was similar to that of the commercial fipronil product. The 20mg/kg dose was selected as the minimum commercial dose rate to provide effective flea and tick control for at least 1 month following a single treatment.

Efficacy of a novel formulation of metaflumizone for the control of fleas (Ctenocephalides felis) on cats.

A novel spot-on formulation containing metaflumizone (ProMeris for Cats, Fort Dodge Animal Health, Overland Park, KS) was evaluated in five laboratory studies to determine the duration of residual efficacy in cats against fleas after a single spot treatment. In each study, eight domestic shorthair cats were randomly allocated to each treatment group and individually housed. One group in each study remained non-treated. In one study, an additional group of eight cats was treated with a placebo formulation. Cats were treated topically with metaflumizone formulation to provide a dose of at least 40mg metaflumizone/kg. Cats were infested with 100 cat fleas (Ctenocephalides felis felis) once per week for approximately 8 weeks. Cats were comb counted 48h after treatment and each infestation to determine the number of viable fleas present. There were no significant differences in flea counts between the non-treated control and the placebo-treated control (P>0.05) other than a 26% reduction at week 1, demonstrating that the formulation excipients had no activity. Metaflumizone treatment resulted in significantly lower flea numbers relative to non-treated controls on all post-treatment count days (P<0.05). Metaflumizone provided >90% control of flea infestations up to 7 weeks following a single treatment.

Confirmation of the efficacy of a novel formulation of metaflumizone plus amitraz for the treatment and control of fleas and ticks on dogs.

A novel spot-on formulation containing metaflumizone plus amitraz (ProMeris/ProMeris Duo for Dogs, Fort Dodge Animal Health, Overland Park, KS) was evaluated in four laboratory studies to confirm efficacy against fleas and ticks on dogs for 1 month. Three different strains of cat flea (Ctenocephalides felis felis) and four tick species were used. Rhipicephalus sanguineus and Dermacentor variabilis were evaluated concurrently in two studies and Ixodes scapularis and Amblyomma americanum in one study each. In all studies, dogs were randomly allocated to treatment groups and compared with nontreated dogs. One study also included a placebo treatment and a commercial product containing fipronil plus S-methoprene. All treatments were applied to the skin at a single spot between the scapulae on Day 0. Dogs were infested with fleas and/or ticks prior to treatment and then reinfested at weekly intervals for 6 weeks after treatment and evaluated for efficacy at 1 or 2 days after treatment and each reinfestation. These studies confirmed that treatment with ProMeris for Dogs at the proposed commercial dose rate rapidly controlled existing infestations of fleas and ticks on dogs. Treatment provided control of reinfesting fleas for up to 6 weeks and at least 4 weeks control of ticks. Efficacy was confirmed in a variety of dog breeds against three different flea strains and four common species of ticks found on dogs in the United States.

Comparative efficiency of flagging and carbon dioxide-baited sticky traps for collecting the lone star tick, Amblyomma americanum (Acarina: Ixodidae).

Collections of Amblyomma americanum (L.) from five habitat types showed that significantly more ticks of all life stages could be collected by CO2-baited sticky traps than by the standard flagging technique. The effect of habitat type and season significantly influenced the number of ticks collected by both techniques. Traps baited with carbon dioxide revealed adult tick activity in several habitats several weeks earlier than did flagging, because flagging failed to efficiently recover ticks unless they had ascended vegetation. The CO2-baited traps required less time and effort and showed less variation for tick population surveys when compared with flagging.

Effect of ivermectin on the volume of blood ingested by two species of ticks (Acari: Ixodidae) feeding on cattle.

Females of the lone star tick, Amblyomma americanum (L.), and the American dog tick, Dermacentor variabilis (Say), were fed on ivermectin (Ivomec)-treated and untreated bovines to determine the effect of the acaricide on volume of blood ingested and to compare the weight differences between the treatment and control groups at various time intervals after attachment. Adult females from each genus were collected from Bos tarus hosts and subjected to hematin assays on three collection dates to estimate the volume of blood ingested. Before feeding, lone star ticks contained an average of 2.0 microliters of blood and had an average weight of 5.2 mg. Unengorged American dog tick females had an average blood volume of 3.3 microliters and a mean weight of 5.8 mg. Ticks of both species reacted to ivermectin by expressing lower mean weights, and they consumed smaller quantities of blood. Lone star tick females were significantly affected in terms of amount of blood consumed and body weight changes when compared with control ticks. After feeding on treated cattle, lone star tick females contained smaller quantities of blood than pretreatment females, but there were no significant differences observed until day 12 between the control and the treated groups. American dog tick females on treated hosts had measurable quantities of blood that were significantly different among the experimental groups.

Persistence of Anaplasma marginale (Rickettsiales: Anaplasmataceae) in male Dermacentor andersoni (Acari: Ixodidae) transferred successively from infected to susceptible calves.

The persistence of Anaplasma marginale Theiler in male Dermacentor andersoni Stiles ticks exposed to the organism as adults was studied as the ticks were successively transferred to five susceptible calves. All calves fed upon by these ticks rapidly developed clinical anaplasmosis; incubation periods of infection ranged from 19 to 26 d and did not change significantly with successive feedings. Development of A. marginale in tick midgut and salivary glands was followed daily during tick feeding (total, 35 d) with light microscopy and DNA hybridization. With microscopy, A. marginale colonies persisted in midgut cells throughout the experiment. Large colonies were observed in gut muscle cells on days 8 through 35 and were the predominant infected cell type during this part of feeding. Colonies were seen in salivary gland acini from day 2 throughout the 35-d experiment. The DNA probe confirmed the presence of Anaplasma DNA in midgut and salivary glands throughout the experiment. Quantitative estimates of infection intensity in tissues of individual ticks approximated 10(7) initial body equivalents, confirming heavy infections. A marginale in midgut tissues decreased with feeding time, whereas the estimated number of organisms in salivary glands remained constant. These data demonstrate that D. andersoni males are efficient vectors of A. marginale and may be potential reservoirs of infection for ruminants for extended periods.

Systemic activity of a benzimidazoline compound in cattle against ticks and biting flies.

A benzimidazoline compound [4-nitro-2-(1,1,2,2-tetrafluoroethyl)-6 -(trifluoromethyl)-1H-benzimidazol-2-, 01, sodium salt] referred to as EL-979 showed systemic acaricidal and insecticidal activity in cattle against 2 tick species, Amblyomma maculatum (Gulf Coast tick) and Dermacentor variabilis (American Dog tick) and adult Stomoxys calcitrans (stable flies). Larvae of black blow fly (Phormia regina) were fed serum collected from treated calves. A complete kill of larvae was obtained with a serum level of 3 parts per million (p.p.m.) of EL-979. Intravenous infusion at the rate of 0.25 mg kg-1 per day for 12 days provided a concentration of EL-979 in the blood of approximately 5 p.p.m. In A. maculatum this blood level prevented feeding in larvae and molting in nymphs, reduced numbers of females successfully feeding and prevented larvae production. D. variabilis adults were not as susceptible to a blood level of 5 p.p.m., but the number of feeding larvae and molting nymphs were reduced. Complete mortality of adult S. calcitrans occurred with repeated feedings of a 5 p.p.m. blood level. Fly control of 80% was found with 2.3 p.p.m. multiple feeds. Calf sera with a concentration range of 16--20 p.p.m. killed all flies which took a single feeding.

Efficacy of selamectin against experimentally induced tick (Rhipicephalus sanguineus and Dermacentor variabilis) infestations on dogs.

Seven controlled studies were conducted to investigate the efficacy of selamectin against weekly infestations of dogs with Rhipicephalus sanguineus and Dermacentor variabilis. Treatments (selamectin or vehicle alone) were applied topically at weekly, 2-week, or monthly intervals or in a "Monthly Plus" regimen (monthly treatment with an additional treatment at 14 days after the first treatment). Selamectin was supplied in unit dose tubes designed to deliver a minimum dosage of 6mgkg(-1). The studies ranged in duration from 37 to 90 days. Fifty adult ticks (+/-2) were applied approximately weekly, and tick counts were performed 3, 4, and 5 days after each infestation. The efficacy of selamectin was expressed as the percentage reduction in geometric mean tick counts on selamectin-treated dogs compared with those for dogs treated with the vehicle alone (negative-control). In one study, the engorgement of Dermacentor variabilis was assessed by weighing ticks after removal on the fifth day after each infestation. Weekly and 2-week interval treatments with selamectin provided efficacies against R. sanguineus of >89% across the entire study periods, with 100% efficacy being achieved from 21 days after the first dose and thereafter (study duration, 37 days for the weekly regimen and 44 days for the 2-week interval regimen). D. variabilis also was well controlled by the 2-week interval treatment regimen, with >96% efficacy being achieved from 21 days after the first treatment and thereafter until the end of the study (study duration: 90 days). In five of six studies incorporating three treatments at monthly intervals, the percentage reduction in R. sanguineus and D. variabilis counts 5 days after infestation ranged from 90 to 100% in the second and third months after treatment began. In the sixth study, reductions of > or =95% in D. variabilis counts 5 days after infestation were achieved for 2 weeks after each treatment in the second and third months. For the Monthly Plus regimen, from the second treatment (day 14) onwards, selamectin achieved 83-100% reductions in R. sanguineus and D. variabilis counts 3 days after infestation, and 94-100% reductions 5 days after infestation in three of the four studies. In the fourth study, selamectin demonstrated good efficacy against D. variabilis for 2 weeks after each treatment. In all seven studies, the counts from the selamectin-treated dogs were significantly (P< or =0.018) lower than those from the vehicle-treated dogs on 77 of the 80 assessments made 5 days after infestation. Selamectin also significantly (P< or =0.0105) reduced engorgement of female D. variabilis. These studies demonstrated that selamectin, administered topically to the skin in a single spot at a minimum dosage of 6mgkg(-1) at monthly intervals, was effective in the control of experimentally induced R. sanguineus and D. variabilis infestations on dogs.

Effect of simulated rain, coat length and exposure to natural climatic conditions on the efficacy of a topical formulation of eprinomectin against endoparasites of cattle.

A series of five controlled studies involving 114 cattle were conducted in Australia, North America and the United Kingdom to examine the effect of simulated rain, coat length and exposure to natural climatic conditions, on the efficacy of a topical formulation of eprinomectin against nematode parasites of cattle. In all trials infections were induced with a range of bovine nematode species and treatment was applied when the majority of nematodes were mature. In one study, simulated rain was applied to cattle ending one hour before treatment or beginning one, three or six hours after treatment. In a second study cattle had short (1 cm) or long (3-6 cm) haircoats at the time of treatment. Three other studies were conducted using cattle housed indoors or exposed to various natural climatic conditions. Nematode counts were determined using standard techniques and the efficacy of treatment was assessed relative to vehicle-treated controls. Regardless of the timing of simulated rain relative to treatment, eprinomectin was at least 99.9% effective (P < 0.01) against Haemonchus placei, Ostertagia ostertagi. Trichostrongylus axei and Cooperia spp. There were also no differences (p > 0.10) in efficacy between treatment administered to dry or wet cattle, or treatment administered before or after simulated rainfall. Efficacies against O. ostertagi, T. axei, Cooperia ancophora and Dictyocaulus viviparus were > 99.5% (p < 0.01) regardless of the length of the haircoat at the application site. Exposure of treated cattle to sunshine and precipitation had no effect on anthelmintic efficacy (p > 0.10) with efficacies of greater than 99.5% being maintained against H. placei, O. ostertagi (adult and fourth-stage larvae), T. axei, Cooperia spp., Nematodirus helvetianus (adult and inhibited fourth-stage larvae) and Oesophagostomum radiatum. These findings indicate that eprinomectin (500 micrograms/kg) in a topical formulation is a safe and highly effective nematocide for cattle regardless of their coat length and this high level of efficacy is maintained in cattle exposed to a wide variety of climatic conditions.

Eprinomectin: a novel avermectin for control of lice in all classes of cattle.

Eight trials were conducted in the United States to determine the efficacy of eprinomectin applied topically against four common species of lice in cattle. In two dose titration trials, eprinomectin dosages of 125 to 750 mcg/kg body weight applied topically were compared to untreated controls. In dose confirmation studies, animals treated topically with eprinomectin applied at the rate of 500 mcg/kg were compared to vehicle-treated controls. Four species of lice were present in these trials: Linognathus vituli, Haematopinus eurysternus, Solenopotes capillatus, all sucking lice, and Damalinia (Bovicola) bovis, the cattle biting louse. Louse counts were made on six to nine predilection sites (the same number of sites in all animals in the same trial) prior to treatment. The same sites were counted again seven days after treatment and weekly thereafter until trial termination eight weeks after treatment. When no lice were found in the predilection sites, a modified whole body search was conducted. Each species of lice was present on at least six animals in each treatment group on at least one counting date in two or more trials. No lice were found on any animal treated topically with eprinomectin at a dosage of > or = 500 mcg/kg after 14 days posttreatment until termination of the trials eight weeks after treatment.

The efficacy of selamectin in the treatment of naturally acquired aural infestations of otodectes cynotis on dogs and cats.

The efficacy of a novel avermectin, selamectin, was evaluated against naturally acquired aural infestations of Otodectes cynotis on dogs and cats. In four controlled and masked studies conducted in the USA and Europe, animals were allocated randomly to treatment with either selamectin at a minimum dosage of 6mgkg(-1) (range, 6-12. 5mgkg(-1)) or the vehicle only from the commercial formulation of selamectin (negative control). Treatments were administered topically in a single spot to the skin of each animal's back at the base of the neck in front of the scapulae. Cats were treated on day 0 only, and dogs were treated either on day 0 only or on days 0 and 30. The ears of dogs were examined otoscopically on day 14 for the presence of viable mites. Mite counts were conducted on day 30 for animals that had received one dose and on day 60 for animals that had received two doses. Percentage reductions in geometric mean mite counts for selamectin treatment compared with the vehicle were 100% for all animals on all count days. Analysis of variance, confirmed by Savage Scores, showed that ln(mite count+1) values were significantly (P< or =0.0015) lower for selamectin than for the vehicle for all animals on all count days. Thus, selamectin administered topically at a minimum dosage of 6mgkg(-1) was safe and 100% effective against naturally acquired aural infestations of O. cynotis in dogs and cats after a single dose or after two doses administered 1 month apart.

Application of fluorescent antibody technique to determine infectivity rates of Amblyomma americanium (L.) (Acarina: Ixodidae) salivary glands and oral secretions by Theileria cervi (Piroplasmorida: Theileriidae).

Salivary glands and oral secretion from individual field collected ticks, or ticks infected with Theileria cervi in the laboratory, were subjected to fluorescent antibody techniques to determine the reliability of the technique in the identification of T. cervi in either the oral secretion or salivary glands. Results indicated that the fluorescent antibody technique can be used with a reasonable degree of success in the identification of infected salivary glands or oral secretion. Oral secretion examinations gave fewer positive samples than when salivary glands from the same ticks were examined.

Effects of Gulf Coast ticks on blood composition and weights of pastured Hereford steers.

Studies conducted in 1975 and 1976 were made of lightweight Herefored steers, pastured on native grass pasture and infested with Gulf Coast ticks, Amblyomma maculatum Koch. The effects of tick infestations on the blood composition and weights of steers over an 8-week period were measured each year and compared to tick-free animals maintained similarly. By the end of the 8-week study infested animals averaged 8.21 kg less than control animals in 1975 and 12.42 kg less in 1976. Of the blood parameters measured significant decreases in Hb, and a lymphopenia, neutrophilia, and basophilia occurred in infested animals in 1975. In 1975 and 1976 after infestations began significant increases occurred in total serum protein, serum globulin, and plasma fibrinogen, and decreases occurred in the albumin/globulin ration of infested animals. Of the serum globulin fractions, only gamma globulin increased in infested animals in 1975 and alpha globulin in 1976. Also, during both years, decreases occurred over time in the number of ticks attaching.

The ultrastructure of sporogony in Theileria cervi (Bettencourt et al., 1907) in salivary glands of female Amblyomma americanum (L.) ticks.

The process of sporogony of Theileria cervi in salivary glands of unfed and feeding Amblyomma americanum females that were infected as nymphs was studied by light and electron microscopy (SEM and TEM). Sporogony was observed only in granular acini type III and in cells classified as complex granular. Infected acini in salivary glands of both unfed and feeding ticks were enlarged. Some stages of parasite development were evident with light microscopy and SEM, but the specific events of sporogony could be documented only with TEM. Nuclear division was evident in unfed adults and continued throughout the developmental process. As the ticks fed, the parasite syncytium became increasingly complex and branches formed that were connected by thin processes. Sporozoites appeared to form by rapid sequential fission of the multinucleate sporont. Initially, sporozoites were oblong in shape and became rounded when mature. Rhoptries and micronemes were located at the hemisphere of the sporozoite that was opposite from the nucleus. Experimentally infected A. americanum transmitted T. cervi to susceptible fawns with a prepatent period of 14 and 20 days. This is the first ultrastructural description of sporogony in T. cervi in tick salivary glands.

Demonstration of Anaplasma marginale Theiler in ticks by tick transmission, animal inoculation, and fluorescent antibody studies.

Cattle were inoculated with a Virginia isolate of Anaplasma marginale Theiler and served as an infective source for laboratory-reared Dermacentor andersoni Stiles and D variabilis (Say) nymphs. Transstadial transmission of A marginale was demonstrated by feeding the newly molted adult ticks of both species on susceptible cattle and by inoculation of gut and salivary gland homogenates collected from adult ticks at postattachment days 5 and 7. Oral secretions collected from the same group of ticks and inoculated into a susceptible cow did not cause infection. Fluorescent antibody studies of infected tick gut and salivary glands were positive, although smears of tick oral secretions were negative for fluorescence.

Ultrastructure of ANaplasma marginale Theiler in Dermacentor andersoni Stiles and Dermacentor variabilis (Say).

Cattle were inoculated with a Virginia isolate of Anaplasma marginale Theiler and served as an infective source for laboratory-reared Dermacentor andersoni Stiles and D variabilis (Say) nymphs. Anaplasma marginale was demonstrated by electron microscopy in gut tissues of replete nymphal ticks and in unfed, incubated, and feeding adult ticks that were exposed to the organism as nymphs when they fed on an infected cow. The A marginale organism in replete nymphs and adult feeding ticks were morphologically similar to A marginale described previously from infected bovine erythrocytes. Colonies of A marginale were demonstrated by light and electron microscopy in midgut epithelial cells of unfed adult D andersoni and D variabilis that had been incubated at 37 C for 3 days. Anaplasma marginale organisms in colonies were very pleomorphic. Small electron-dense particles were demonstrated in all infected tick stages studied, but were most evident in colonies from incubated, unfed ticks. They may represent a reproductive form of A marginale.

Morphologic characteristics of colonies of Anaplasma marginale Theiler in midgut epithelial cells of Dermacentor andersoni Stiles.

Colonies of Anaplasma marginale Theiler were studied in midgut epithelial cells of adult Dermacentor andersoni Stiles that were infected as nymphs. Colonies were categorized by light microscopy into 5 morphologic types that appear to represent stages in a developmental sequence. The mean diameter of the colony types ranged from 5.64 micrometer to type 1 colonies to 10.49 micrometers of type 5 colonies. Representatives of each colony type were selected by light microscopy and were sectioned for examination by electron microscopy. The morphologic features of individual A marginale organisms within colony types varied and included: (i) small electron-dense forms, (ii) larger reticulated forms, (iii) pleomorphic reticulated forms, and (iv) small particles. Types 1 and 2 colonies contained small, electron-dense forms. Type 3 colonies contained electron-dense forms, early reticulated forms, and small particles that were often outside of the parasite's limiting membrane. Type 4 colonies contained many reticulated forms that often had small particles within cell membranes. Type 5 colonies contained fewer well-formed reticulated forms than did type 4 and, in some cases, large masses of pleomorphic reticulated forms.

Longevity of colonies of Anaplasma marginale in midgut epithelial cells of Dermacentor andersoni.

Colonies of Anaplasma marginale in midgut epithelial cells of experimentally infected Dermacentor andersoni were studied in adult ticks 1, 3, and 6 months old. Longevity of the parasite in ticks was assessed by evaluating its infectivity for splenectomized calves; calves were exposed by feeding ticks and by inoculation of tick gut homogenates. Longevity was also evaluated by determining size, type, and density of colonies in male and female ticks. The effect of incubation (2.5 days at 37 C) on colony density was also examined for ticks at each age period. All methods used to assess longevity of A marginale in ticks (tick transmission, calf inoculation, and histologic studies) indicated a decrease of the numbers of organisms in 6-month-old ticks. Furthermore, when tick gut homogenates from 6-month-old nonincubated ticks were not infectious for susceptible calves, incubation of ticks before dissection restored infectivity of homogenates. Colonies of A marginale were detected in gut tissues of 6-month-old ticks that were not infective; therefore, infectivity of ticks could not be confirmed merely by presence of A marginale colonies.

Percutaneous infection of nymphal Dermacentor andersoni with Anaplasma marginale.

Newly replete nymphal Dermacentor andersoni (principals) were percutaneously exposed to Anaplasma marginale by injection of either intact or lysed infected bovine erythrocytes. Control nymphs were fed on calves with anaplasmosis. The subsequently molted adults were examined for infection by light microscopy, and companion ticks were tested for infectivity by allowing them to feed on susceptible calves. When they fed as adults, both control ticks and percutaneously inoculated principals transmitted A marginale to susceptible calves. Prepatent periods in calves varied according to the method by which nymphs were infected. Colonies of A marginale were found in all ticks that acquired infection by feeding, but colonies were not observed in any ticks exposed percutaneously. The possible developmental cycle of A marginale in artificially infected ticks is discussed.

Preliminary studies of the development of Anaplasma marginale in salivary glands of adult, feeding Dermacentor andersoni ticks.

On each day of feeding on susceptible calves, salivary glands obtained from groups of adult ticks that transmitted Anaplasma marginale were examined for A marginale colonies by use of light microscopy and transmission electron microscopy. On day 8 of feeding, salivary glands were examined, using fluorescein-labeled antibody and methyl green-pyronine stain. Use of fluorescein-labeled antibody consistently revealed small numbers of fluorescent foci in salivary gland acinar cells obtained from ticks that had fed for 8 days. Colonies of A marginale were seen by transmission electron microscopy only in salivary gland acini of male ticks; these colonies could not be identified, using light microscopy, in companion 1-micron plastic sections stained with Mallory stain. Methyl green-pyronine stain, used commonly to detect theilerial parasites in tick salivary glands, did not differentiate A marginale from cytoplasmic inclusions normally found in salivary gland acinar cells.