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Prevalence of Dog Erythrocyte Antigen 1 in 7,414 Dogs in Italy.

1. Introduction

Canine blood groups are recognized and have standardized terminology as Dog Erythrocyte Antigen (DEA). The DEA system includes seven well-known blood groups (DEA 1, 3, 4, 5, 6, 7, and 8) with over twenty antigen specificities [1-4]. Other nonstandardized antigens within DEA such as Dal [3, 5] and the recently described Kai 1 and 2 have been reported [6]. Canine blood groups comprise a two-allele system with a positive and negative type.

The DEA 1 blood group system initially was described with 3 types, DEA 1.1, 1.2, and likely 1.3 [3, 7, 8]. Utilizing quantitative flow cytometry and an immunochromatographic technique with a monoclonal anti-DEA 1 alloantibody the continuum of DEA 1 negative to weakly (1+) up to strongly DEA 1 positive (4+) blood type was observed in contrast to the originally described DEA 1 system and with a significant correlation between these tests [9]. Recently, an autosomal dominant mode of inheritance of 4 alleles of DEA 1 with strong (4+) to weak (1+) reactivity was discovered. DEA 1 positive alleles are dominant over DEA 1 negative allele without any direct correlation with the historical DEA 1.2 positive subtype [10]. So now, it is accepted to speak about the DEA 1 group as a whole without any subtype [9,10].

Natural occurring antibodies against DEA 1 antigen in dog's erythrocytes have never been positively identified [11]. On the other hand, natural antibodies against DEA 3, 5, and 7 have been documented with a prevalence of 6%, 23%, and 45%, respectively [2]. However, although all canine blood group antigens can stimulate the formation of alloantibodies, DEA 1 seems to be the most immunogenic and also is considered the most clinically important. Alloantibodies will appear following the first transfusion in DEA 1 negative recipient dogs receiving positive DEA 1 red blood cells (RBCs) within 4-14 days [11-14]. Sensitization of the recipient and production of alloantibodies can result in a severe acute hemolytic transfusion reaction and even death if a second DEA 1 positive RBC transfusion is administered to the same patient [15-17]. The risk of alloantibody production and transfusion reactions against antigens other than DEA 1 is not yet well defined [2] and there is no documented clinical evidence of a hemolytic reaction caused by DEA 1.2, 3, 5, and 7 in mismatched transfusions [18].

Blood typing to identify the presence of DEA 1 and the cross-match to establish full compatibility should be performed before each transfusion in order to reduce the risk of sensitization or immunological reaction between donor and recipient dogs [2, 19, 20]. Available methods for typing DEA 1.1 antigen include agglutination cards (RapidVet-H[R], Agrolabo, Scarmagno, TO, Italy) and an immunochromatographic strip for DEA 1 (Quick Test DEA 1[R], Alvedia, Lyon, France) both using monoclonal antibodies and are useful for in-clinic testing [17,21]. A gel column agglutination test using microtubes is generally reserved for use in laboratory settings. The advantage of gel column typing is the ability to establish the degree of agglutination that can be graded from 0 to 4+. A DEA 1.1 positive result is considered if the reaction is graded as [greater than or equal to] 2+ [17,18, 22].

The knowledge of breed differences for prevalence of DEA 1 is very important for the recruitment of typed compatible blood donors. DEA 1 is expressed approximately in 40-60% of the general canine population. The prevalence of canine blood group DEA 1 has been studied in small populations in geographically restricted areas [23-28]. The most frequently studied breeds were German Shepherd, Golden Retriever, Greyhound, Doberman, and Rottweiler along with investigations in mongrel dogs [4,18, 28-30].

The aim of our study was to determine the prevalence of blood type DEA 1 in a large population database of purebred and mongrel dogs reared in Italy. Furthermore, we calculated the potential risk of sensitization following a first transfusion and the subsequent risk of an acute transfusional reaction as documented immune-mediated hemolysis and/or agglutination following the second transfusion in the absence of a pretransfusional cross-match and blood typing test.

2. Material and Methods

The population of this study included 7,414 dogs retrieved from the Italian database of the website Dog blood Donors (DbD) (http://www.dogblooddonors.it). The data collected were recorded in the website during the period of October 2014-July 2016. The following information has been extracted from the DbD database: date of the dog's registration on the website, breed, weight, sex, year of birth, age, type of blood group (negative or positive DEA 1), and the Italian regions where the dog was living at the time of registration. Dogs were enrolled by private practitioners who recorded the above data into the DbD website.

DEA 1 blood type was established using two commercial in-clinic typing tests: RapidVet-H, Canine DEA 1.1, Agrolabo (Scarmagno, TO, Italy), and Quick TEST DEA 1 Alvedia (Lyon, France, distributed in Italy by Alcyon Italy, Marene, CN). Both tests use a monoclonal antibody, the former in an agglutination assay and the latter in immunochromatographic strips. Dogs were typed using one of the two in-clinic tests. Nevertheless, the RapidVet-H Canine DEA 1.1 test was the one most used in up to 95% of the dogs.

The data collected from the website DbD has been organized as follows:

(i) The prevalence of DEA 1 negative and positive types in the total population and in breeds that showed a substantial number of subjects registered (over 50) was calculated.

(ii) The breeds numbering over 50 subjects was sub-typed as females and males and the prevalence of DEA 1 was calculated according to sex.

(iii) In the breeds numbering over 50 subjects the percentage of the breed's population was determined in relation to the total Italian canine population inferred from the last 10 years of registration at the ENCI (Ente Nazionale di Cinofilia Italiana, Italian National Kennel Club) (http://www.enci.it) which was calculated.

(iv) The data concerning the provenience and corresponding blood type of the 7,414 dogs was subdivided into different geographical regions of Italy.

(v) Using the standard inclusion criteria for canine blood donors (dogs heavier than 25 kg and aged between 2 and 8 years) and breeds numbering over 20 subjects were considered as prospective "blood donors" regardless of whether they were DEA 1 positive or negative.

(vi) The risk of sensitization for each breed (over 50 subjects) was calculated using the formula [(% DEA 1 negative x % DEA 1 positive)/100] to establish the probability of a dog to become sensitized after the first transfusion of blood without having been tested with a cross-match and typed for DEA 1 [28, 30, 31].

(vii) Using the following formula [(% DEA 1 negative x % DEA 1 positive) x (% sensitization for the first transfusion/10,000)] the probability of each breed (over 50 subjects) to develop an acute hemolysis and/or agglutination (immune-mediated) transfusional reaction with a second incompatible transfusion using uncross-matched and untyped blood was calculated [28, 30, 31].

(viii) The Excel[R] 2016 software (Microsoft Office) was used for data analysis.

3. Results

In Table 1, the prevalence of DEA 1 negative and positive types in the canine population registered in the database of the DbD website in Italy (breeds represented by over 50 dogs) is reported. The prevalence of DEA 1 negative and positive types in breeds with less than 50 subjects and the complete list of breeds are not shown.

In Table 2, the prevalence of DEA 1 negative and positive types for females and males for breeds represented by over 50 dogs is reported.

In Table 3, the percentage of breed's population in relation to the total Italian canine population inferred from the last 10 years of registration at the ENCI for the breeds numbering over 50 subjects is reported.

In Table 4, the prevalence of prospective blood donors (PBD), using inclusion standard criteria for canine blood donors (including dogs heavier than 25 kgs and aged between 2 and 8 years) is reported. Only breeds with more than 20 subjects were considered.

In Table 5 the risk of sensitization following the first transfusion and the risk of an acute transfusional reaction documented as hemolysis and/or agglutination (immune-mediated) following the second transfusion in the absence of pretransfusional cross-match and blood typing test according to the most involved breeds were shown.

In Figure 1 the distribution of DEA 1 negative and positive types in the several Italians regions is shown.

4. Discussion

This paper reports the largest study among the references available on DEA 1 blood group prevalence to date. In addition, is also includes the DEA 1 prevalence in breeds never reported before. Indeed, a high number of purebred canine subjects have been tested here, and the results provide substantial information about the prevalence of DEA 1 blood group in Italy, which could be useful not only in transfusion medicine but also for canine genetic epidemiologic studies.

To obtain the most reliable results about the prevalence of DEA 1 in the canine purebred population in Italy, only breeds represented by over 50 dogs were analyzed. The prevalence of PBD in purebred and mixed dog's breeds represented by more than 20 subjects also was examined. Of total 7,414 dogs tested (purebreds 4,798, mongrel 2,616) 2,878 (38.8%) were DEA 1 negative and 4,536 (61.2%) were DEA 1 positive (Table 1). The prevalence for DEA1 positive dogs found in this studywas very similar to results 553/890 (62.0%) reported in another study carried out in Italy [29]. Furthermore, there have been other studies that showed similar results for DEA 1 positive dogs as in the case of Nigeria 71/178 (60.1%), Turkey 116/178 (65.2%), Portugal 156/274 (56.9%), and Spain 110/206 (53.4%) [4, 25, 28, 32]. Nevertheless, the first research about blood groups distribution in dogs from 1961 reported a lower prevalence of DEA 1 positive dogs 148/332 (44.6%) [13]. The difference here could reflect the effects of the ongoing worldwide inbreeding and linebreeding practiced within the more popular dog breeds over the ensuing years. A recent study in eastern Africa reported as much as 78/100 (78.0%) for DEA 1 positive dogs [33], representing the highest reported prevalence for DEA 1 positive blood types. Unfortunately, the paper did not report which breeds were included and the canine population studied was quite small. It is well known that the DEA 1 blood group distribution varies according to the geographical area investigated [6].

The DEA 1 blood group in dogs has been studied mainly for transfusional purposes. Transfusion reactions usually do not occur following the first transfusion because naturally occurring antibodies against DEA 1 are very rarely found in dogs (<0.3% in a population of 2,500 dogs tested by Hale and Werfelmann) [34], unless they have been previously sensitized by an incompatible transfusion. Regardless of this, DEA 1 negative dogs should receive only DEA 1 negative blood. If a negative DEA 1 dog is transfused with DEA 1 positive erythrocytes, an antibody reaction (agglutination and/or hemolysis) can be elicited in the recipient [2].

The good clinical practice in canine transfusion medicine requires that both recipient and donor should be blood typed for DEA 1, which is considered highly antigenic and the most clinically relevant blood type [2, 35]. In addition, the cross-match test is advisable to be carried out before a transfusion. If a recipient receives consecutives transfusions of incompatible blood, this patient could be highly sensitized and develop a severe acute hemolytic reaction [15,16].

In this study a high prevalence for DEA 1 positive type in breeds such as Rottweiler (89.5%), Golden Retriever (75.2%), and Dachshund (74.2%) was reported. These results are comparable to those found in other previous investigations: 78.0-100% in Rottweiler, 77.0-95.0% in Golden Retriever, and 71.0-100% in Dachshund [24, 28, 29, 36, 37]. It is important to know the frequency of DEA 1 positive blood type in the Rottweiler and Golden Retriever as they could be enrolled as blood donors, because their weight is in the range established for donations and they are generally docile dogs. The high prevalence for breeds such as Ariegeois (95.7%), English Setter (81.0%), and Zwergpinscher (80.0%) has been confirmed according to the previous study carried out in Italy [29].

In this study, the Labrador Retriever breed showed a high prevalence for DEA 1 positive type (65.3%) as reported previously in studies carried out in Switzerland, South Africa, and Italy [24, 29, 37]. Nevertheless, in the published study from Portugal, Labrador Retrievers showed a higher prevalence for DEA 1 negative type (55.0%), although those results could be related to the low number of dogs (number 29) tested in comparison to our data (number 478) [28].

The prevalence of DEA 1 positive type in English Cocker Spaniel is 65.2%, in the Poodle 68.8%, and in Yorkshire Terrier 63.0%. These values are different from those found in other countries such as Switzerland which reported a high prevalence for DEA 1 negative (67%) type in the Poodle and in Portugal which reported 68.8% of DEA 1 negative type in the English Cocker Spaniel [28, 37].

In this study, the Shih Tzu breed showed a high prevalence for DEA 1 positive type (72.2%), while, on the contrary, a Japanese study reported an almost equal prevalence for DEA 1 positive (57%) and negative (43%) types in this breed. It should be pointed out, however, that only a small number of dogs (7) were tested in the Japanese study [38].

A high frequency for DEA 1 positive blood type has been observed in breeds such as French Brittany Spaniel (77.1%), Pug (77.0%), English Springer Spaniel (74.1%), and Chihuahua (61.0%). These results cannot be compared due to the absence of previous studies (Table 1).

In this study breeds with high DEA 1 negative prevalence were German Shepherd (81.1%), Boxer (83.0%), French Bulldog (77.9%), and West Highland White Terrier (63.0%) (Table 1). The high prevalence for DEA 1 negative type in German Shepherd was similar to that found previously in few countries such as South Africa (84.0%) [24], Portugal (100%) [28], Brazil, and Italy (both 90.0%) [29, 36]. Almost the same situation was observed in the Boxer, in addition to South Africa (88.0%) [24], Portugal (100%) [28], Switzerland (100%) [37], and Italy (80.0%) [29]. It is noteworthy that DEA 1 negative frequencies in French Bulldog (77.9%) and West Highland White Terrier (63.0%) were not previously reported. A high prevalence of DEA 1 negative type could be expected from the mating between female and male dogs within the same blood line but it is unknown if these tested dogs were the result of inbreeding or linebreeding. A recent study showed that mating between DEA 1 negative dogs strictly produced DEA 1 negative offspring, while mating between DEA 1 positive dogs primarily resulted in DEA 1 positive offspring, with an occasional DEA 1 negative offspring [10].

In our report, several breeds such as Jack Russell Terrier, Beagle, Border Collie, and Maltese showed an almost equal prevalence of DEA 1 negative and positive subjects. On the other hand, in this study, the Cane Corso breed shows exactly 50.0% for both DEA 1 negative and positive subjects, and these results are completely different from those obtained in another study carried out in Italy (72.0% for DEA negative type) [29]. The DEA 1 negative type prevalence reported in South Africa for American Staffordshire Terrier was 75.0% which differs from the equal prevalence found in the current study [24]. Another breed whose prevalence differs from those found in this study is the Maltese breed which reported a 33% for DEA 1 positive type in a study carried out on only 9 Maltese dogs in Japan [38]. The prevalence found in Cavalier King Charles Spaniel (48.5% of DEA 1 positive and 51.5% of DEA 1 negative types) dogs cannot be compared with any previous study as far as we know (Table 1).

Differences in the DEA 1 frequency in various countries are generally due to the selection of the population tested and the genetic drift deriving from local breeding. It is important to consider that in this study the sampling for DEA 1 testing was using dogs randomly accessed from several veterinary facilities and privately owned. However, as the blood type is a genetic marker, it should be considered as one way to reveal the hybrid history of canine breeds supplying information about migration and breed development in the canine database of several countries. Of course, this information is less accurate than the genomic data in the phylogenetic analysis reported by Parker et al. (2017) [39].

Breeds such as French Brittany Spaniel, Pug, English Springer Spaniel, and Chihuahua had a higher prevalence for DEA 1 positive type. On the other hand, breeds that had higher prevalence for DEA 1 negative type were French Bulldog and West Highland White Terrier. So far, there are no studies that report the DEA 1 prevalence for these breeds, although it would be interesting to be able to compare these results, even though we are not sure if the dogs included in this study from matings between inbred or tightly linebred dogs. Perhaps the fact that some of these dogs presented a high percentage of positive or negative DEA 1 blood type could be the result of matings between two DEA 1 negative dogs that produced only DEA 1 negative offspring or matings between DEA 1 positive dogs that resulted in DEA 1+ dogs, with an occasional production of a DEA 1 negative dog [10].

There was no notable difference in the prevalence for DEA 1 negative or positive type among females and males in this study, except for females Boxers (13/18 DEA 1 positive dogs were females), which represented 72.2%. The Cavalier King Charles Spaniel (35/50 DEA 1 positive dogs were males) and West Highland White Terrier (14/20 DEA1 positive dogs were males) both represented 70%. The Ariegeois breed females had the highest DEA 1 negative (100%) type but only 4 females were tested (Table 2). It would be interesting to know if Ariegeois negative bitches had received blood transfusions so they could become sensitized. In this occurrence, the mating with a DEA 1 positive dog could raise the probability of risk of neonatal isoerythrolysis in any DEA 1 positive offspring [2].

There are not any studies about the prevalence of DEA 1 negative or positive types in relation to sex and breed except an investigation carried out in East Africa showing generic similar result but in a very small population [33]. A previous study established that the DEA 1 blood group system is an autosomal trait, with both male and female dogs either being DEA 1 negative or having varying degrees of DEA 1 positivity [10]. In addition, a survey ascertained that female dogs, even after pregnancy, did not develop any alloantibodies against RBC antigens during gestation, so they can be used safely as blood donors [40]. Further, additional pretransfusion compatibility testing is not required should they require transfusions themselves.

The obtained percentages of purebred dogs tested for DEA 1 in comparison to the numbers of these breeds enrolled by ENCI in the last 10 years were more than 1.0% in Yorkshire Terrier, Ariegeois, Shih Tzu, Maltese, and Cavalier King Charles Spaniel, and a high consistency (almost 3.0%) was noted in the Zwergpinscher breed (Table 3). These percentages provide important genetic epidemiologic data for the prevalence of DEA 1 in Italian purebred dogs.

The data compiled in the DbD website permitted us to achieve the aims of the project in the several Italian regions covered (Figure 1). The northern Italian regions were apparently more aware of the need for blood donation in dogs in comparison to the other Italian regions. This situation seems to be the same in humans blood donors where a higher percentage of donors were in regions such as Emilia Romagna (3.2%), Marche (3.5%), and Lombardia (2.6%) [41]. In addition, the low number of enrolled dogs in southern Italian regions could also be related to the low number of dogs officially registered with a microchip, since this data was a mandatory requirement to enroll dogs in the DbD website.

The so-called "universal donor" dogs are generally accepted as being negative for DEA 1,3,5, and 7 but positive only for DEA 4. Usually DEA 3, 5, and 7 blood groups are not tested because they do not show a major transfusion reaction during the first transfusion and there are difficulties in obtaining the required blood typing antisera [2].

Anyhow, the prevalence of alloantibodies against blood group antigens in dogs is quite rare and the only alloantibody consistently found is against DEA 7 antigen (around 10% in 2,500 dogs tested), and so far evidence of transfusion reactions or neonatal isoerythrolysis has not been documented [17, 34, 40].

In this study, 852/1745 (48.8%) of the purebred dogs were considered as PBD. A smaller number of PBD have been found in the mixed breed dogs (304/2616, 13.9%) (Table 4). The status of PBD was established using the standard inclusion criteria for canine blood donors as data reported in the DbD web page, namely, dogs being heavier than 25 kgs and of ages between 2 and 8 years, regardless of whether they were DEA 1 positive or negative and female or male (Table 4). Obviously, due to their selection as PBD dogs the distribution of DEA 1 negative and positive type is different from the general population studied and reported in Table 1.

Most of the breeds in this study have the sensitization risk ranging from 20.0 to 25.0% which means that the probability of a dog to become sensitized following the first transfusion of blood without having been tested with a cross-match and typed for DEA 1 is quite high (Table 5). Rottweiler and Ariegeois breeds had only a minor risk to become sensitized and produce antibodies against DEA 1 following the first transfusion (9.4 and 4.2%, resp.). These results are related to the higher prevalence of DEA 1 positive type in these breeds which reduced the risk of transfusional reactions. On the other hand, Beagle, Cane Corso, and Cavalier King Charles Spaniel had the highest percentage likelihood to become sensitized (25.0%) (Table 5).

Regarding the risk of an acute transfusional reaction documented as hemolysis and/or agglutination following the second transfusion, Ariegeois and Rottweiler breed showed the minor risk (0.2-0.9%). On the contrary Cane Corso, Beagle, Cavalier King Charles Spaniel, American Staffordshire Terrier, Border Collie, Jack Russell Terrier, and Maltese showed the highest risk (6.3 to 6.1%) (Table 5). Only a few studies have reported the risk for sensitization of an acute transfusional reaction. In 4 native breeds of Turkey a high sensitization risk after the first transfusion (14.4-35.6%) and the risk of an acute hemolytic reaction after second transfusion (7.2-25.3%) were found [32]. In Ibizan Hound and Galgos from Spain the sensitization risk after the first transfusion was similar to the data reported here [4, 30].

A limitation of this study was the use of the two in-clinic blood typing methods. Misclassification of blood typing results could occur even if performed by an experienced person [21]. Indeed, taking pictures of the results and storing them for future needs are suggested, but we were not able to have access to these raw data. Considering the high number of animals tested, however, even if a few errors in blood typing classification occurred, they would not have influenced the overall conclusions of the data presented. Finally, when used in healthy dogs both assays gave almost equivalent results [21].

Regardless of this, at present there is no a gold standard for blood typing in dogs [16]. The Quick TEST DEA 1 Alvedia and the RapidVet-H Canine DEA 1.1 are easy to use and were performed by veterinary practitioners. Nevertheless, the proper use and interpretation of the rapid typing test RapidVet-H Canine DEA 1.1, which was largely used in this study, were completed upon request by the Agrolabo, TO, Italy technical support group [8,21, 22,42].

5. Conclusions

This study provides an overview about the distribution of DEA 1 blood group in a large dog population belonging to purebred and mongrel dogs reared in Italy. The data could be useful in clinical transfusion medicine and for studies of canine genetic epidemiology. The prevalence of DEA 1 positive and negative dogs mostly agrees with previous prevalence studies reported in the literature. In addition, DEA 1 distribution was studied in breeds never previously reported. The risk of sensitization following the first transfusion and the risk of an acute transfusional reaction documented as hemolysis and/or agglutination following the second transfusion in the absence of a pretransfusional cross-match and blood typing test also was determined in most of the breeds studied.

Disclosure

Part of the data of this paper was presented as abstract at the 56th AIVPA Annual Conference, 5-7 May, 2017, Piacenza, Italy.

Conflicts of Interest

The authors declare that there are no conflicts of interest regarding the publication of this paper.

https://doi.org/10.1155/2017/5914629

Acknowledgments

The authors wish to thank Dr. W. Jean Dodds of Hemopet, Garden Grove, California, USA (http://www.hemopet.org), for her review of the manuscript. The authors acknowledge the Almo Nature SpA that created the DbD project from which the data was extrapolated.

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[32] E. Ergul Ekiz, M. Arslan, M. Ozcan et al., "Frequency of dog erythrocyte antigen 1.1 in 4 breeds native to different areas in Turkey," Veterinary Clinical Pathology, vol. 40, no. 4, pp. 518-523, 2011.

[33] S. Dhliwayo, T. A. Makonese, B. Whittall, S. M. Chikerema, D. M. Pfukenyi, and M. T. Tivapasi, "A study on the prevalence of dog erythrocyte antigen 1.1 and detection of canine babesia by polymerase chain reaction from apparently healthy dogs in a selected rural community in zimbabwe," Journal of the South African Veterinary Association, vol. 87, no. 1, Article ID a1409, 2016.

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[42] B. Kohn, G. Classe, and C. Weingart, "Clinical evaluation of the QuickVet[R]/RapidVet[R] canine dog erythrocyte antigen 1.1 blood-typing test," Journal of Veterinary Diagnostic Investigation, vol. 24, no. 3, pp. 539-545, 2012.

Anyela Andrea Medina Valentin, Alessandra Gavazza, and George Lubas

Veterinary Transfusional Center (CTV), Department of Veterinary Sciences, University of Pisa, Via Livornese Lato Monte,

San Piero a Grado, 56122 Pisa, Italy

Correspondence should be addressed to Anyela Andrea Medina Valentin; anyela.medina@vet.unipi.it

Received 11 May 2017; Revised 30 June 2017; Accepted 1 August 2017; Published 24 September 2017

Academic Editor: Ingo Nolte
Figure 1: Distribution of DEA 1 negative and positive dogs in
Italians regions (according to raw data retrieved from http://
www.dog-blooddonors.it website).

Regions          Dog tested
                    No.

Lombardia          3,697
Piemonte           1,025
Emilia Romagna      948
Toscana             493
Umbria              388
Veneto              330
Marche              145
Lazio               139
Liguria             111
Sardegna             64
Puglia               34
Sicilia              17
Other regions        23
Total              7,414

TABLE 1: Prevalence of DEA 1 negative and positive in the breeds.

Breeds                           Dogs        DEA1            DEA
                                 tested       neg           1 pos
                                  No.       No. (%)        No. (%)

Mixed                            2,616     924 (35.3)    1,692 (64.7)
Labrador Retriever                478      166 (34.7)     312 (65.3)
German Shepherd                   312      253 (81.1)     59 (18.9)
Golden Retriever                  214      53 (24.8)      161 (75.2)
Jack Russell Terrier              213      100 (46.9)     113 (53.1)
Zwergpinscher                     194      62 (32.0)      132 (68.0)
Chihuahua                         164      64 (39.0)      100 (61.0)
English Cocker Spaniel            158      55 (34.8)      103 (65.2)
American Staffordshire            153      72 (53.1)      81 (46.9)
  Terrier
Dachshund                         149      35 (25.8)      114 (74.2)
Poodle                            144      45 (31.3)      99 (68.8)
Maltese                           130      59 (45.4)      71 (54.6)
English Setter                    126      24 (19.0)      102 (81.0)
Border Collie                     124      66 (53.2)      58 (46.8)
Beagle                            123      61 (49.6)      62 (50.4)
Yorkshire Terrier                 108      40 (37.0)      68 (63.0)
Boxer                             106      88 (83.0)      18 (17.0)
Cavalier King Charles Spaniel     103      53 (51.5)      50 (48.5)
Ariegeois                          92       4 (4.3)       88 (95.7)
French Bulldog                     86      67 (77.9)      19 (22.1)
French Brittany Spaniel            83      19 (22.9)       64 (771)
Shih Tzu                           79      22 (27.8)      57 (72.2)
Cane Corso                         76      38 (50.0)      38 (50.0)
Pug                                61      14 (23.0)       47 (770)
Rottweiler                         57       6 (10.5)      51 (89.5)
English Springer Spaniel           54      14 (25.9)      40 (74.1)
West Highland White Terrier        54      34 (63.0)      20 (37.0)
Other *                          1,159     440 (38.0)     719 (62.0)
Total                            7,414    2,878 (38.8)   4,536 (61.2)

* Breed represented by less than 50 dogs.neg, negative, pos,
positive, No., total number, and %, percentage (according to raw data
retrieved from http://www. .dogblooddonors.it website).

TABLE 2: Prevalence of DEA1 negative and positive in female and
male dogs.

                            Dogs                     DEA1 neg
Breeds                      tested     Females         Males
                             No.       No. (%)        No. (%)

Mixed                       2,616     475 (51.4)     449 (48.6)
Labrador Retriever           478      80 (48.2)      86 (51.8)
German Shepherd              312      120 (47.4)     133 (52.6)
Golden Retriever             214      33 (62.3)      20 (37.7)
Jack Russell Terrier         213      4 8 (41.0)     59 (59.0)
Zwergpinscher                194      28 (45.2)      34 (54.8)
Chihuahua                    164      32 (50.0)      32 (50.0)
English Cocker Spaniel       158      28 (50.9)      27 (49.1)
American Staffordshire       153      37 (51.4)      35 (48.6)
  Terrier
Dachshund                    149      21 (60.0)      14 (40.0)
Poodle                       144       22(48.9)      23 (51.1)
Maltese                      130      33 (55.9)      26 (44.1)
English Setter               126      12 (50.0)      12 (50.0)
Border Collie                124      37 (56.1)      29 (43.9)
Beagle                       123      30 (49.2)      31 (50.8)
Yorkshire Terrier            108      20 (50.0)      20 (50.0)
Boxer                        106      40 (45.5)      48 (54.5)
Cavalier King                103      20 (37.7)      33 (62.3)
  Charles Spaniel
Ariegeois                     92      4 (100.0)       0 (0.0)
French Bulldog                86      29 (43.3)      38 (56.7)
French Brittany Spaniel       83      13 (68.4)       6 (31.6)
Shih Tzu                      79       8 (36.4)      14 (63.6)
Cane Corso                    76      15 (39.5)      23 (60.5)
Pug                           61       6 (42.9)       8 (57.1)
Rottweiler                    57       3 (50.0)       3 (50.0)
English Springer Spaniel      54       7 (50.0)       7 (50.0)
West Highland White           54       16 (471)      18 (52.9)
  Terrier
Other *                     1,159     219 (49.8)     221 (50.2)
Total                       7,414    1,429 (19.3)   1,449 (19.5)

                                             DEA 1 pos
Breeds                        Females         Males
                              No. (%)        No. (%)

Mixed                        895 (52.9)     797 (47.1)
Labrador Retriever           160 (51.3)     152 (48.7)
German Shepherd              25 (42.4)      34 (57.6)
Golden Retriever             85 (52.8)      76 (47.2)
Jack Russell Terrier         54 (47.8)      59 (52.2)
Zwergpinscher                66 (50.0)      66 (50.0)
Chihuahua                    46 (46.0)      54 (54.0)
English Cocker Spaniel       52 (50.5)      51 (49.5)
American Staffordshire       37 (45.7)      44 (54.3)
  Terrier
Dachshund                    58 (50.9)      56 (49.1)
Poodle                       40 (40.4)      59 (59.6)
Maltese                      39 (54.9)      32 (45.1)
English Setter               38 (37.3)      64 (62.7)
Border Collie                32 (55.2)      26 (44.8)
Beagle                       25 (40.3)      37 (59.7)
Yorkshire Terrier            36 (52.9)      32 (47.1)
Boxer                        13 (72.2)       5 (278)
Cavalier King                15 (30.0)      35 (70.0)
  Charles Spaniel
Ariegeois                    42 (47.7)      46 (52.3)
French Bulldog                8 (42.1)      11 (57.9)
French Brittany Spaniel      26 (41.9)      38 (61.3)
Shih Tzu                     22 (38.6)      35 (61.4)
Cane Corso                    22 (579)      16 (42.1)
Pug                          19 (40.4)      28 (59.6)
Rottweiler                   28 (54.9)      23 (45.1)
English Springer Spaniel     17 (42.5)       23 (575)
West Highland White           6 (30.0)      14 (70.0)
  Terrier
Other *                      353 (49.1)     366 (50.9)
Total                       2,259 (30.5)   2,277 (30.7)

* Breed represented by less than 50 dogs; neg, negative, pos,
positive, No., total number, %, percentage (according to raw data
retrieved from http://www.dog-blooddonors.it web site).

TABLE 3: Breeds in relation to the overall size of dog breed
population according to the ENCI last 10 years of registration.

Breeds                            No. of     ENCI      %
                                   dogs      data
                                  tested

Zwergpinscher                       194      6,732    2.88
Yorkshire Terrier                   108      7,664    1.41
Ariegeois                           92       7,174    1.28
Shih Tzu                            79       6,620    1.19
Maltese                             130     12,816    1.01
Cavalier King Charles Spaniel       103     10,291    1.00
Poodle                              144     15,185    0.95
French Bulldog                      86       9,551    0.90
English Cocker Spaniel              158     20,528    0.77
West Highland White Terrier         54       7,297    0.74
Beagle                              123     17,407    0.71
Pug                                 61       9,099    0.67
American Staffordshire Terrier      153     23,011    0.66
Labrador Retriever                  478     77,116    0.62
Border Collie                       124     23,101    0.54
Dachshund                           149     28,108    0.53
Chihuahua                           164     32,494    0.50
Jack Russell Terrier                213     46,221    0.46
Golden Retriever                    214     49,580    0.43
Boxer                               106     36,926    0.29
English Springer Spaniel            54      20,473    0.26
Cane Corso                          76      31,296    0.24
German Shepherd                     312     152,649   0.20
Rottweiler                          57      30,350    0.19
French Brittany Spaniel             83      44,534    0.19
English Setter                      126     145,906   0.09

ENCI: Italian National Kennel Club, No.: total number, %, percentage
(according to raw data retrieved from
http://www.dogblooddonors.itwebsite).

TABLE 4: Number of prospective blood donors.

Breeds                            DEA 1 neg     DEA1 pos    * PBD
                           No.     No. (%)      No. (%)      No.

Labrador Retriever         478    166 (34.7)   312 (65.3)    264
German Shepherd            312    253 (81.1)   59 (18.9)     171
Golden Retriever           214    53 (24.8)    161 (75.2)    133
American Staffordshire     153    72 (53.1)    81 (46.9)     58
  Terrier
Boxer                      106    88 (83.0)    18 (17.0)     66
Ariegeois                  92      4 (4.3)     88 (95.7)      5
Cane Corso                 76     38 (50.0)    38 (50.0)     27
Rottweiler                 57      6 (10.5)    51 (89.5)     32
Bernese Mountain Dog       44      8 (18.2)    36 (81.8)     33
Dobermann                  32     25 (78.1)     7 (21.9)     19
Kurzhaar                   30      7 (23.3)    23 (76.7)      4
Australian Shepherd        29      9 (31.0)    20 (69.0)      3
Siberian Husky             29     12 (41.4)     17 58.6)      4
Maremma Sheepdog           24     12 (50.0)    12 (50.0)     15
Weimaraner                 24     14 (58.3)    10 (41.7)     13
Belgian Shepherd           23     12 (52.2)     11 (478)      3
English Pointer            22      8 (36.4)    14 (63.6)      2
Total                     1,745   787 (45.1)   958 (54.9)    852

Breeds                     DEA neg      DEA pos
                           No. (%)      No. (%)

Labrador Retriever        105 (39.8)   159 (60.2)
German Shepherd           145 (84.8)   26 (15.2)
Golden Retriever          38 (28.6)    95 (71.4)
American Staffordshire    24 (41.4)    34 (58.6)
  Terrier
Boxer                     56 (84.8)    10 (15.2)
Ariegeois                  1 (20.0)     4 (80.0)
Cane Corso                17 (63.0)    10 (37.0)
Rottweiler                 3 (9.4)     29 (90.6)
Bernese Mountain Dog       7 (21.2)    26 (78.8)
Dobermann                 14 (73.7)     5 (26.3)
Kurzhaar                   2 (50.0)     2 (50.0)
Australian Shepherd         0(0.0)     3 (100.0)
Siberian Husky             2 (50.0)     2 (50.0)
Maremma Sheepdog           7 (46.7)     8 (53.3)
Weimaraner                 7 (53.8)     6 (46.2)
Belgian Shepherd           2 (66.7)     1 (33.3)
English Pointer            1 (50.0)     1 (50.0)
Total                     431 (50.6)   421 (49.4)

* PBD prospective blood donor based on age (2 to 8 years old) and
body weight (>25 kg); neg, negative, pos, positive, No., total
number, and %, percentage (according to raw data retrieved from
http://www.dogblooddonors.it website).

TABLE 5: The risk of sensitization after 1st transfusion and the
risk of an acute hemolytic reaction after 2nd transfusion in the
absence of pretransfusional cross-match and blood typing
test ([section]).

                                 Sensitization         Acute
Breeds                           risk after 1st       immune-
                                  transfusion        mediated
                                                     reaction
                                      (%)            after 2nd
                                                  transfusion (%)

Beagle                                25.0              6.2
Cane Corso                            25.0              6.3
Cavalier King Charles                 25.0              6.2
Spaniel
American                              24.9              6.2
Staffordshire Terrier
Border Collie                         24.9              6.2
Jack Russell Terrier                  24.9              6.2
Maltese                               24.8              6.1
Chihuahua                             23.8              5.7
West Highland White                   23.3              5.4
Terrier
Yorkshire Terrier                     23.3              5.4
Mixed                                 22.8              5.2
English Cocker                        22.7              5.1
Spaniel
Labrador Retriever                    22.7              5.1
Zwergpinscher                         21.7              4.7
Poodle                                21.5              4.6
Shih Tzu                              20.1              4.0
English Springer                      19.2              3.7
Spaniel
Dachshund                             19.1              3.7
Golden Retriever                      18.6              3.5
French Brittany                       17.7              3.1
Spaniel
Pug                                   17.7              3.1
French Bulldog                        17.2              3.0
English Setter                        15.4              2.4
German Shepherd                       15.3              2.4
Boxer                                 14.1              2.0
Rottweiler                            9.4               0.9
Ariegeois                             4.2               0.2

([section]) According to raw data retrieved from
http://www.dogblooddonors.it website.
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Title Annotation:Research Article
Author:Valentin, Anyela Andrea Medina; Gavazza, Alessandra; Lubas, George
Publication:Veterinary Medicine International
Date:Jan 1, 2017
Words:7654
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