15900941 virullence in goats

7/27/2019 15900941 Virullence in Goats

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Materials and MethodsAnimals Fifty-four Boer-Spanish crossbred weanling

goats were used in the* study. Goats were 10 to 16 weeks old. They were housed in a 3-sided bam in 9 pens (6 goats/pen);

each pen was 7,420 cm4. Goats were administered medica

tions to eliminate internal parasites' and coccid ia Goatswere liiitfed a com mercial {ielletized ration (44% grata con

centrate, 20 % alfalfa hay, 30% cotton seed hulls and meal, 5%(potasses, vitamins A and E, and trace minerals): water wau

available ad libitum. The experimental protocol wasapproved by a regional animal care and use committee.

Experimental design-Goats were allowed to acclimate to thei r en vironment for 2 weeks. Each pen of goats was then randomly allotted to 7 treatment, groups and J control

groups. The tent control group was 'used to evaluate the effectof being confined in a tent filled with aerosolized dust for 4

hou rs followed In intrat racheal administration of 30 ml oisaline (JS.9% MaCl) solution; Goats in the pen control group

Were no t man ipulated tie, no t Confined in the dust-fil led tent stud did not receive intratracheal inoculations):.

Administration o f aerosolized dust to goats Goats inthe 7 treatment groups and tent, control group were exposed to aerosolized fcedyard dust. The feedvard dust preparation fas been th..ictciL.ed and described elsewhere ' Stee ol dustparticles was determined by dry dispersion. Three 1-g sam

ples were each passed through an instrument to determinedistribution of mean particle sire. The dust was aatodaved3

for 15 minutes at 1 21 C and 6.9 kPa of pressure to killmicrobes (bacteria, fungi, and fungal spores) typically foundin feedyard dust.* Cultures of autodaved dust did not yield

viable colonies of bacteria or fungi.The aerosolization process has been described in detail

elsewh ere.1,1* Briefly, d ust was administered to g oats con tained in a semi-airtight tent.' A 1,5 00 g aliquot of sterilized

feedyard dust was placed in a hopper that angered the dust into a funnel, which led to a jet mill* Dissemination of dust by the jet mill was augmented with air produced by a gaso-

line-pQwered portable air compressor* and air from a largeblower motor.1Throughout a period of 4 hours, goats were

exposed to 950 g of aerosolized dust inside the tent. The 4- hour dust treatment preceded intratracheal administration oifungal spores (treatm ent grou ps) o r saline solution (tent, control group). Goats were exposed to aerosolized dust once

weekly for 6 consecutive weeks.

Preparatio n of fungal spo res State .of the ? fungal sporetypes was as follows: Aspergillus fumigcUusand PtMiaUium

ch tysogmum, 2 to 5 pm; Trichodermavir k k , 3 to 4 pm:

Chm hv nk m g iobost imand M ucor mmmi smmus, 4 to. 8: jsa;

Stachyb& t rys cha t tH mm *5 to 9 |tm: anil Mimotospora lanu-

ginosa,>8: to lO jin i.S ix of these fungi arc commonly found infeedyard dust,"' with the e xception being S e k a n m m , whichis commonly found on grain. All 7 types, of fungi used in thestudy were identified by personnel at a microbiological labo-ratory1that specialized in fungi identification.

Each species of fungus was incubated on 30 malt-extract

agar1plates (10 0 X 15 m m) at 28G for sufficient time (5 to14 days) to enable development of massive sanita tion ,. EachPetri plate* was washed with 5 ml. of sterile physiologic saline solution, and contents of the 30 plates were pooled. Sterile saline solution was added to achieve a final volume of 210 m i. T o separate spores from hyphae, the fungal suspen

sion was; mixed and strained twice through a 30-ro.ra, No,140 sieve," Concentration of spores in the suspension was coun ted irt a hem acytom eter.' The sp ore stispensiBns: were;

also titrated in triplicate on mak-extract agar Petri plates to calculate the mean number of viable CPUs pet milliliter.

Antibacterial inhibitors {cM ortet racy cline / 5 mg/L; streptomycin sulfate/1 10 0 mg/L) were incorporated into malt-

extract agar to prevent bacterial growth. Each spore preparation was mixed and divided equally into six 30-mL. syringes

labeled appropriately for each group of fungi, Weekly for 6weeks, each spore type was inoculated immediately aftergoats were expos ed to aerosolized dust for 4 hours.

Intratracheal inoculation of fungal spores *Contentsof the syringes were mixed thoroughly immediately prior to tracheal inoculation. Initially, goats in 4 treatment groups

(A fum i ga tm , P ch t ymgmum, M ranwsissimus, and A! lanuginosa)and the tent control group were administered an anes

thetic locally at the site of the needle insertion for the intra- mu heal inoculation, ant! a sedative (xylazine hyd rochloride')was administered 1M. Thirty'Milliliters of the spore prepara

tion (or saline solution for the tent control group) was inoculated into the lumen of the proximal portion of the trachea

at a location just distal to the crico id cartilage. However, sev

eral of these goals died as a result of Idiopathic reactionsrelated to the, sedative.

Therefore, goats in the remaining 3 treatment groups (C gfobmim , T viridc,and Schnrtaram) were anesthetized forfungal inoculation. Goats were administered atropine sulfate*(0.04 mg/kg, 1M), which was followed by an IV injection of a combination of hutorphanol tartrate0 (0.5 mg/kg) and ket-amine hvdroi Monde >'4.4 mg/kg; A specul um* with a light

source was inserted into the orai pharynx of each goat, and

the epiglottis was viewed. Then a 1-tnL pipette* equippedwith a tip consisting of a hollow rubber tube was inserted

past the epiglottis into the proximal portion of the trachea. Ange: containing the fungal spore solution was attached topipette, and the 30 ml. of spore preparation was inocu

lated into the trachea.

latte 1Mean S6M number of total, viable, .and rusnvtabtefun! ^r e s/ 3 E H rl . dose asniirtisteradweeWy to each groups of goats (6 goats/group) during a 6-week pariod.

Viable Nonviabls spares Total

AspergiHusfumipatus

Chastomiumglobosum

Monotosporlanuginosa

Myeor

Periieilliumchrysogemsm

Stachyfaotr/schartarum

Tricliotirtnawiritfe

0.495 X 10s 0.381 XI 0

0,7038 X 10s 0.2566 X Itf

0.030 X 10s i 0.010 X10 5

0.115 X tO8 0.689 X 109

0.214 x 10s 0.026 * 10'*

1,287X10 0.804 X 10*

4.494X 10* 1.597 X 10*

2.377 .X10 1.065 X 10s

4.649 X 10* 1,964X10

7.068X10*+. l'5B3X 10"

0.041 X 10 0.02702 X 10* 0,751 10 ' 0.2644X10 0.7920 105 0.2894 X10

7.831 X W s i G.4414X 10 5.247 X 1 # i 1.418X10* 6 H I X f ! i 1.126 X1 0"

1.782 X 10 1.052X10

5.198X10* 1.509X10*

2.408 X 10s 1.066X10*

4.764 - 10 r 1.996 X 10

7.282 X 10 1.554 X 10*

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Collection o f data and samples Prior to each inoeula-tkm of fungal spores, blood samples were collected from a jugular vein and rectal temperature# were obtained fromeach; goat. Reel#! temperatures were obtained 0, 4, 24, 48,

and 72 hours after tnoailatioiv of fiwsgal spores, and blood

samples were collected for WBC counts 24 and 72 hours afterinoculation of fungal spores.

Necrop sy At. completion of the study , all goats were tobe euthanatized by IV admia ktratio n o f art overdose of a barbiturate solution. One goat from each group was euthanatized 24 hours after the sixth inoculation of fungal, spores,and the remaining goats tn each group were euthanatized 72; hoa rs after the last spore inoculation;. All goats were necrop-

Sied by a veterinary pathologist (J'RA), Gross and histologic examinations of tissues were performed by the pathologist,,

who was no t aware of the group status for each goat.During necropsy, sterile cotton-tipp ed applicators* were

used to collect mucus and tissue fluid


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\

when they were removed front the tent after a 4hourexposure. The internal rtares were covered with thedust, which became moistened by vapor from respira-tion. Some goals had intermittent coughing after they

were removed from the tent; however, this coughing

Figure 2 Mean * SEM total WBC counts for a tent controlgroup !A; solid circles) and each of 4 spore-treated groups(A fumiggtus (open squares). M lanuginosa (open triangles!,M ramosissimus {open diamonds! and P chrysogenum (openhexagons!) and 3 pen control group (8; solid circles) arid each of the 3 remaining spore-treated groups (C. globoaum (opensquares). S chartarum [open triangles], and T vi'ride [open diamonds!! of goats. S ee Figure 1 for remainder of key

subsided within 24 hours. Intratracheal inoculations ofthe 30ml. suspension of fungal spores were more dif-ficult than had been anticipated. The needle slippedout of the lumen of the trachea in a few instances.

Muco r ranw sissimm spores were prone to induce anabscess w hen they were deposited outside the lum en ofthe trachea. Clinically, M ramosissimus and T virideappeared to be m ore pathogen ic than the other types offungi. This clinical impression of these 2 fungal typeswas reinforced by the increase in reeial temperatureand by gross and histologic lung lesions.

We detected significant (P < 0,001) differences inmean rectal temperatures and mean WBC counts amonggroups following inoculation of the various fungal spores(Figu res I and 2). Mean rectal temperature of each of thespore treatment groups for the first 4 treatment groups(ie, A ju m ig atm . P chrysogenum , M ramosissimus, aridM lanuginosa) was compared with mean rectal tempera-ture of the tent control group. For goats inoculated withM ram osiss imu s, mean rectal temperature was signifycandy increased in weeks 1 through 4 and week 6 at 24,48, and 72 hours after inoculation. The exception to thispattern was week 5, when the mean rectal temperaturedecreased at 72 hours. For goats inoculated withP chrysagamm spores, mean rectal temperature signifi-cantly increased in week 2 (24 and 48 hours after inocu-lation) and week 6 (72 hours after inoculation). Forgoats inoculated with M lanuginosa spores, mean rectaltemperature was significantly increased in week 4 (72hours after inoculation). Mean rectal temperature forgoats inoculated with fum igatu s was significantlyincreased in week 5 at 24 and 48 hours after inoculation.

Mean rectal temperature of the remaining 3 sporetreatment groups (ie, Cglob osum , t vir ide , and S char-tarum) was compared with that of the pen control goats.

Goats that received T viride spores had a significantincrease in mean rectal temperature in week 1 (24 and48 hours after inoculation), weeks 2 and 3 (24 hoursafter inoculation), weeks 4 and 5 (4 hours after inocu-lation), and week 6 (4 and 24 hours after inoculation).For the C gtohosum group, mean rectal temperature wassignificantly increased in weeks I and 2 (24 and 48hours after inoculation), week 3 (4, 24, and 48 hoursafter inoculation), week 4 (4. 24, 48, and 72 hours after

Table- 2 Mean SO size of atelectatic lesions, in each group of goats *

Right lung_________________________________le ft fung

Fungal type Severity! No. of goats Cranial Middle Caudal Cranial Middle Caudal

M ramosissimus* n 4 4,700 1,536 2,650 1,109 5,745 - 4.171 1,350 1,559 2,325 862 2,200 2,540

T viride 21 6 3,318 i 3,859 1,560 1,695 2,364 2,312 7,849 Z 12,593 1,833 1,066 851 - 636S chartarumS 3* 6 2,767 t 900 1,993 :i 670 3,133 1,876 2,053 1,440 1,656 t 756 1,201 832C giobosumS 41 6 2,781 z 1,086 r^

s$ 1

400 1,321 1,262 386 665 990 532 190 S 425

P chtysogenum* 5 6 2,425 1,611 1,750 ,t 920 500 i 1,225 200 489 333 : 532 1,650 t 4,042A fumigates*' 6 5 2,142 1,279 1,710 971 150 * 141 1,350 1.559 2,325 862 2,200 2,540M lanuginosa* 7 8 906 * 1,001 581 1,102 0 0 0 0Tent control* 8 6 1,133 1 489 88 161 50 r 123 33 82 75 184 0Pen control 9 6 649 r 1,166 420 * 698 0 0 97 124 0

Values reported are tire number of square millimeters for each lesion, which was obtained by multiplying the width and length of each lesion.Each treatment group was exposed to aerosolized sterile dust for 4 hours followed by intratracheal inoculation with 30 ml. of a fungal spore

suspension weekly for 6 weeks, the tent control group was exposed to aerosolized sterile dust for 4 hours followed by intratracheal injection with30 mL of physiologic saline 10.3% NaClj solution weekly for 6 weeks; and the pen control group was not exposes to aerosol ized ster ile dust anddid not receive intratracheal injections; f Severity represents the ranking {greatest severity of lesions to least severity of lesions! and was deter-mined by adding values for lesions in all lung lobes in that group. tValues differ significantly (P s 0.05; Dunnatt paired f test) from values for thetent control group. IValues for this treatment group were compared with values for the pen control group.

: ( Week t | Week 2 Week 3 W&ek 4 W ee k S W eek 6

r ** & * w V*1tst *

o y* 2Ck 3 | Week 4 WeekS J

O * f*


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inoculation), week 5 (4, 24, and 48 hours after inocu-lation}, and week 6 (4 and 24 hours after inoculation).M em rectal temperature was significantly increased forthe S chartarum group in week 2 (24 and 48 hours after

inoculation), week 3 (4, 24, and 72 hours after inocu-lation), week 4 (4, 24, and 48 hours after inoculation),week 5 (4, 24, and 48 hours after inoculation), andweek 6 (4 and 72 hours after inoculation).

Mean WBC count of the initial 4 treatment groupswas compared with the mean WBC count of the tentcontrol goats. For goats inoculated with P chiysogenum,mean WB C cou nt was significantly increased in week 1(24 hours after inoculation) and week 4 (72 hours afterinoculation). F or goats inoculated withM ram oshs im us,mean WBC count was significantly increased in week 1(24 and 72 hours after inoculation) and weeks 3 and 5(24 hours after inoculation). Mean W BC c ount was sig-nificantly increased for goats inoculated with A fu mig a is in week 5 at 24 and 72 hours after inoculation.

Mean WBC count of the remaining 3 treatment groupswas compared with the mean WBC count of the pen con-trol goats. For goats inoculated with S chartamm, meanWBC count was increased in weeks 1 through > (24 hoursafter inoculation).. For goats inoculated with C g l ok m m ,the mean WBC count was increased in weeks 1 and 4 (24hours after inoculation), weeks 2 and 6 (72 hours afterinoculation), and weeks 3 and 5 (24 and 72 hours afterinoculation). The mean WBC count was increased for goatsinoculated with T ytride in. weeks I and 5 (24 hours afterinoculation) and week 2 (0 hours [lime of inoculation]).

The thermophilic fungus M lanuginosa appeared tobe the least pathogenic of the fungal types. However, 1goat inoculated with M lanuginosa died: during the studyas a result of bronchopneumonia complicated by rumen

acidosis.Necropsy Atelectasis was the most common

lesion observed during necropsy (Table 2). Atelectasiswas most prominently observed in goats inoculatedwith M ftimostssimuSL Consolidated lung lesions were,also observed in goats inoculated with M ramasmimus,T vtride, S chartarum , and C globosum.

Histologic examination of respiratory tract tissuesrevealed that goats from the tent control and M lanuginosa groups had mild, patchy atelectasis with bronchialassoci-

ated lymphoid tissue. Fungal spores and mi tall fragmentsof hyphae were observed in some sections of the goatsinoculated with M lanuginosa, and no spores or hyphaewere observed histologically in lung sections of the tent

control group. Tissues from goats inoculated with M r a m o stssimws had an airway exudate consisting of prdteinaceousfluid that contained many macrophages and neutrophils,with a few granulomas; some lung sections containedmuhinucleated giant cells. There was patchy fibrosis in .anumber of histologic lung sections from goats inoculatedwith M ramosissimus, Aj'umigatus, and P chrysogcnum.These groups also had patchy to coalescing airway exudatethat contained macrophages, neutrophils, and giant, cells.There was a noticeable increase in periairwayassociatedlymphoid tissue in these 3 fungal groups, and fungalspores' and small fragments of hyphae were observed insome sections. Substantial microscopic lung lesions wereobserved in the pen control group. Goats inoculated withCgtobosutn. S chartarum, and T viiidc had patchy to gen-

eralized alveolar and airway exudate that containedmacrophages and neutrophils, with granulomas and a fewgiant cells in some sections. Fungal spores and small frag-ments of hyphae were, observed in macrophages in most ofthe sections examined.

Use of Goinori stain revealed that all lung sectionsfrom negativecomrol goats were negative for fungalelements and that all sections from goats in the treat-ment. groups were positive for fungal elements inmononuclear cells. Fungal spores in lung sections ofgoats inoculated with M mmosissimui or Afumigatus had buds and germinal tubules.

Subjectively, severity of lesions differed among thevarious fungal organisms inoculated imo the lungs ofgoats, in desce nding order of severity o f lesion, the

fungal organisms were M mm asmimu s, T viride, C glo-bosum, 5 chartarum, A fumigatus, P chrysogenum, andM lan ugino sa.

Diagnoses were established for infections resultingfrom the various fungal organisms. Mucor ramosissimus caused chronic exudative mycotic pneumonia with granu-loma formation, wftereas A fum iga tus and P chtysogcnumcaused chronic exudative pneumonia. Inoculation ofC globosum, S chartamm, and T yiride caused chronicactive pyogranulomatous pneumonia.

table 3 Mean SD number of fungal isolates cultured from various tissues obtained from goats in each treatment group.

No. of goats

Bronchial Thora cic Proximal aspect Distal aspectFungal type 24 hours* 72hours* lymph fluid of trachea of trachea Bronchi Lungs

A fumigatus It 5 0 0 4.0 8.8 3.0 7.4 0 261 t 285C globosum t 5 0.7 >


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Isolation of fungi from respiratory tractsVarious amounts of viable spores were frequentlyrecovered from the respiratory tract of goats et all 7treatment groups (Table 3). Isolation of fungi from thenasal cavity was not reported because it frequently con-tains environmental fungi of all types. The bronchiallymph nodes and thoracic fluid of all goats inoculatedwith T viride yielded fungal spores.

DiscussionEnvironmental fungi can induce disease in domestic

animats and humans via infections onaor in a host"'*8byinducing.allergic reactions initiated bv inhalation of deadar living fungus spores or fragments o f hyphae,27* andthrough production of toxins. Toxins are most oftenintroduced into a host through ingestion of contaminat-ed foods. fa humans, it has been proposed* that toxiceffects can be induced by inhalation of fungal toxins;however, this hypothesis is controversial.

When small dust particles of low toxicity fill alveo-

lar macrophages deep in the lungs, the phagocytes maybe overloaded and impaired. One hypothesis is thatoverloading impairment is caused by suppressed pul-monary clearance and there is a concurrent recruitmentof alveolar macrophages and neutrophils in the alveolararea.1 In addition to overloading, macrophages may alsobe damaged by toxins, mycotoxins, and proteolyticenzymes from the phagocytized dust and microbes,uThere is an increase in the number of particles that enterthe interstitial space. The percentage volume of parti-cles may be more important than the numbe r of particlesin overbading of alveolar macrophages,* Pulmonaryclearance of ultrafine particles is much slower than forlarger particles.* There may be a prolonged inflammato-ry response with persistent and excessive elaboration of

chemotatic substances and an elaboration of proinflammatory cytokine s.11 This may allow fungal spores toescape phagocytosis. It is also conceivable that fungalspores could infect pr release immunosuppressive mycotoxinsif l that may damage the delicate lung tissues, Thepossible overloading of alveolar macrophages with ster-ilized feedyard dust, may decrease the immune responsesufficiently to allow inoculated fungal spores to inducetissue damage prior to their elimination from the lungs.The endotoxin in organic manure dust activatesmacrophages through CD 14 receptor sites.^ Host pul-monary defenses to Rhizopus and Aspergil lus vary evenin clinically normal animals. Host defenses against

Aspergillus spores depend on early killing of the sporesby alveolar macrophages, whereas the prevention of

mucormycosis appears to require that bronchioalveolarmacrophages inhibit the conidia from converting intothe hyphal form; however, these macrophages areunable to kill Rhizopus oryzue spores.0

The study reported here revealed which, of the 7types of spores was the most toxic when infused 6times into the trachea of nonimmunosuppressed goats.Histologic examination of lung tissues collected fromeach of the 7 groups revealed that the fungal elements(spores and small fragments of hyphae) were evident24 and 72 ho urs after inoculation of spores into the tra-chea. It has been reported that viable P chrysogenumspores were not cultured from the lungs of mice when

samples were obtained > .36 hours after exposure.Spore buds and germination tubules were seen twice inI. goat from each o f theA fumigatus and M ra mps iM mus groups.

Size of the fungal spores ranged front 2 to 7 jim for6 of the 7 fungal groups and was 8 to 16 pm for M lanu-ginosa. It would appear that the upper end of the rangeof the spore sizes could not possibly reach the alveoli inhumans; however, it appears that, some spores: of all 7groups reached the alveoli: in goats. Mo ntospara lanuginow had the largest spore size and also had the leastseverity of observed lung lesions (not significantly dif-ferent from the severity for the control groups).Nevertheless, M lanuginosa spores wi?re observed in thealveolar region. Perhaps there is a wider radge of sporesizes for this fungi, especially smaller spores, than hasbeen published. Certainly, viable spores were isolatedfrom the lung tissues of all 7 groups of goals.

Of the 7 groups of fungi tested, the most invasiveappeared to be Tviriik as determined on the basis of its

isolation in greatest numbers from the thoracic fluid andthoracic lymph nodes of all goats in that group.Triehoderma organisms were isolated from both lungs in5 of 6 goats, the exception being that organisms werenot isolated from the middle and caudal lobes of the left,lung of 1 goat. A few fungal CPUs were isolated from thebronchial lymph nodes of at. least 1 goat that was inocu-lated with C globosum and S c h a n a n m , and no fungiwere isolated from the thoracic fluid of any of the 7groups. Mucor mm osis siinus was extremely pathogenicwhen inadverten tly deposited into the soft tissues of theneck, and it caused large abscesses that probably con-tributed to the deaths of 2 goats. Mucor organisms inoc-ulated via the trachea also induced severe damage to theventral half of all 3 lobes of the right lung in 4 goats and

to all 3 lobes of the left lung in 2 of those goats.The tnycotoxin for Penicillium and Aspergillus

(excluding afiatoxms that induce aflatoxicosis in birdsand farm animals that eat contaminated food) appears tobe of low toxicity,*5 Gliotoxin and helvolic acid report-edly4' are produced byA Jumiga ius . Pen id llimn verruco surn and Aspergil lus oehm ceu s produce the mycotoxinochrato xin A,4i We were not able to find any informationon m ycotoxin production byM onotospom organisms.

Four treatment groups (M ranmsissimus, T viritfc,S chartantm, and C globosum) had significantly largeratelectatic and consolidated lung lesions, compared withlesions for the other 3 treatment groups or the 2 controlgroups. Excluding M ramosissimus, 3 of thesfc fungalgroups produce m yc oto xirts.^ * Stachyb&tiys char

iarum, an important fungus with a worldwide distribu-tion, produces macrocydic triebothecenes referred to assatratoxins G and H'1 that are potent inhibitors of pro-tein and DNA synthesis,51 It grows well in plaster wallboards,51 ceiling tiles, and particle boards from housesthat have been damaged by water leaking through roofsor water pipes. It is frequently called black mold by theinsurance industry, and most, if not all, insurance com-panies no longer cover this mold infestation in theircommon homeowner policies because of the number ofclaims that were made. People who remain in houseswith this mold are at risk of serious health deterioration,probably through inhalation1 of fungal elements that:

62 0 AJVR, Vol 66, NO, 4, April 2Q0S


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contain the mycatoxin.. This fungus is believed, but notproven, to induce infant idiopathic pulmonary hemor-rhage syndrom e:.^ Trkhodemui produces trichotheceneihycotoxms,8 which a:re frequently referred to as trieho

dermol. and its acetyl ester uichoderarin. Chaeiamium globosu m produces the highly toxic chaetomins andchaetoglobosins mycotoxins.5

The exception appears to be M ramosissimus, Wewere not able to find anything on mycotoxin productionby this organism in the literature, One publication* onZygomycetes in humans indicated that there are 2 ordersof Zygomycetes (Mttcorales and Entvmophthmiles) andthat most of the illnesses were caused by Mucoraks.

Mu cora les includes Rhizopus spp. Other fungi associatedwith human illness were Mucoi, Rhiztm ucor, Abs idia , and5 other fungal types. Infections with M uc orak s organ-isms commonly lead to thrombosis and infarctions ofinvolved tissues and result in tissue destruction that: isoften mediated by a number of fungal proteases, lipases,anti mycotoxins. The spores from these furtgi are trans-mitted by inhalation or ingestion.5* Infection with Mucprspp results in an extremely aggressive mucormycosis inimmunocompromised humans* and can also induceinfections in humans that are not immunocompro-mised.* Mucormycosis has been diagnosed in many ani-mal species, including marine mammals,1 cattle,sheep,'1 swine,M hor ses,* platypus es* tree frogs,* andbirds.68

Spores from 4 types of fungi (M ramosissimus, T viritfc, C glohosum, and 5 chartamm) induced substantialpathologic changes in the lungs when healthy goatsreceived repeated intratracheal inoculations, It is suspect-ed that the pathologic changes, atelectasis, and consoli-dated lesions were caused by fungal exoproducts (ie, proteases, lipases, and mycotoxins). These substances were

probably released when spores were destroyed by the hostmacrophages and neutrophils; alternatively; they couldhave been actively produced by the spores. Additionalinvestigations will be necessary to provide answers. Theatelectatic lung lesions observed in the tent control goatswere induced by exposure to aerosolized sterile dust priorto intratracheal injectio n of 30 mL of sterilized physiolog-ic saline solution. Similar lesions were not seen in the pencontrol goats that were not exposed to aerosolized dust oradministered saline solution via intratracheal injections.

a. Ivoraec. MSD AGVET, Merck & Cafae, Rahway, .Nj.b. Amproiktm, MSD AGVET, Merck & Co inc. Rahway, NJ.c. Malvern Mastersizer 200 0, version '5=1, Malvern: Iissfnmfaits

Ltd, Malvern, UK.d. Autoclave, Amseo 2021 eagle series, Steris Corp, Healthcare

Division, Erie, Pa.e. Custom-fabricated canvas Ktit, Wolfe Canvas, Amarillo, Tex.t AeC.uRate


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